Reviews

The principles of directed cell migration

Shuvasree SenGupta1, Carole A. Parent1,2,3,4 and James E. Bear 5,6 ✉

Abstract | Cells have the ability to respond to various types of environmental cues, and in many
cases these cues induce directed cell migration towards or away from these signals. How cells
sense these cues and how they transmit that information to the cytoskeletal machinery governing
cell translocation is one of the oldest and most challenging problems in biology. Chemotaxis,
or migration towards diffusible chemical cues, has been studied for more than a century, but
information is just now beginning to emerge about how cells respond to other cues, such as substrate-​
associated cues during haptotaxis (chemical cues on the surface), durotaxis (mechanical
substrate compliance) and topotaxis (geometric features of substrate). Here we propose four
common principles, or pillars, that underlie all forms of directed migration. First, a signal must be
generated, a process that in physiological environments is much more nuanced than early studies
suggested. Second, the signal must be sensed, sometimes by cell surface receptors, but also in
ways that are not entirely clear, such as in the case of mechanical cues. Third, the signal has to be
transmitted from the sensing modules to the machinery that executes the actual movement, a
step that often requires amplification. Fourth, the signal has to be converted into the application
of asymmetric force relative to the substrate, which involves mostly the cytoskeleton, but perhaps
other players as well. Use of these four pillars has allowed us to compare some of the similarities
between different types of directed migration, but also to highlight the remarkable diversity in
the mechanisms that cells use to respond to different cues provided by their environment.

1
Department of Pharmacology,
University of Michigan Medical
School, Ann Arbor, MI, USA.
2
Department of Cell and
Developmental Biology,
University of Michigan Medical
School, Ann Arbor, MI, USA.
3
Rogel Cancer Center,
University of Michigan,
Ann Arbor, MI, USA.
4
Life Sciences Institute,
University of Michigan,
Ann Arbor, MI, USA.
5
UNC Lineberger
Comprehensive Cancer Center,
University of North Carolina
at Chapel Hill School of
Medicine, Chapel Hill,
NC, USA.
6
Department of Cell Biology
and Physiology, University
of North Carolina at Chapel
Hill School of Medicine,
Chapel Hill, NC, USA.
✉e-​mail:
jbear@email.unc.edu
https://doi.org/10.1038/
s41580-021-00366-6
Cell migration is critical in a wide array of physiological,
developmental and disease-​related processes, and basic
tenets governing this process have been uncovered over
the years. In vivo, cells must be able to perceive a vari-
ety of cues in their environment and migrate towards or
away from these cues so as to execute morphogenetic
programmes during development, mount an immune
response and repair damaged tissues. When this process
goes awry, devastating consequences often ensue. Failure
of cells to migrate in the appropriate way can lead to
defects during neuronal development linked to cognitive
deficits1, chronic wounds that never heal2 and immune
deficiencies3. Improperly initiated or misdirected cell
migration can be equally detrimental, leading to invasive
metastatic cancer4, autoimmune disease5 and fibrosis6.
Biologists have studied the process of directed migra-
tion for more than a century, but many mysteries remain
about how this process works at a mechanistic level.
To define how cells move directionally towards var-
ious cues, it is critical to understand the basics of cell
migration. Perhaps the most influential paradigm for
describing cell migration is the four-​step cycle of cell
crawling developed by Michael Abercrombie, an early
pioneer of the field7. This paradigm arose from his
observations of migrating primary and cultured fibro-
blasts by phase contrast and interference reflection
microscopy8. In this scheme, the first event is the protru-
sion of a leading edge, which in fibroblasts is dominated
by lamellipodia and filopodia. Next, the cell generates
initial adhesions with the substrate. These adhesions
connect to the contractile machinery of actomyosin
stress fibres and, through a combination of pulling from
the front and squeezing from the rear, the cell body
moves forward. Finally, old adhesions are detached from
the substrate or dissolved at the trailing edge. Although
textbooks describe this as a stepwise cycle of sequential
steps, in reality, all events of the cycle occur simultane-
ously and likely influence the other steps rather than
proceeding as a series of independent events. In addi-
tion, many of the concepts of the Abercrombie cycle
are specific to mesenchymal cells such as fibroblasts on
2D surfaces and may not apply to other cell types that
use different modes of migration or during migration
in different physiological environments (see Box 1 for a
primer on different modes of migration).
In this Review, we present a four-​part conceptual
framework for understanding directed cell migration
towards a variety of cues, including diffusible chemical
cues (chemotaxis), chemical cues on a surface (hapto­
taxis), mechanical substrate compliance (durotaxis),
geometric features of the substrate (topotaxis; also known
as contact guidance) and electric fields (galvanotaxis;

NAture RevIews | MolECular CEll BioloGy volume 22 | August 2021 | 529

0123456789();:
Reviews
Box 1 | Modes of cell migration and plasticity
Cells can migrate singly or collectively as groups227–229. Historically, single-​cell migration
has been divided into mesenchymal and amoeboid modes of migration, although
these classifications are complicated by the plasticity of migration modes in different
environments (see below). Fibroblasts, various stem cells and some cancer cells often
use the mesenchymal migration mode, which is defined by several characteristics, such
as a strong dependence on adhesion to the extracellular matrix (ECM), an elongated
morphology in 3D environments, actin-​based protrusions such as lamellipodia or
filopodia at their leading edge, and the ability to generate strong traction forces on the
substrate through contractile actin networks. These characteristics usually give rise to
slower migration velocity93. The amoeboid migration mode is used by a wide range of
cells, including primordial germ cells, single-​cell social amoebas such as Dictyostelium
discoideum and immune cells such as leukocytes230. In this mode, cells exhibit a more
rounded morphology, undergo constant shape changes through rapid extension and
retraction of membrane protrusions, and engage with the substrate through weak
adhesions, which usually lead to higher migration velocity. Amoeboid protrusions
vary from lamellipodia and filopodia driven by actin polymerization to actin-​free
transient spherical blebs that rely on myosin-​based contraction and pressure-​driven
cytosolic flow217,230.
Cells can also move collectively as groups, which presents its own challenges and
opportunities. Both epithelial and mesenchymal cells exhibit collective migration,
which is important for tissue remodelling during morphogenesis, wound closure and
cancer cell invasion211,229. During collective migration, signals from external cues are
transmitted to the entire mass of cells through the integration of intracellular and
intercellular signalling cascades as well as mechanotransduction at cell–cell junctions
and cell–ECM interfaces212. This results in front–rear polarity at a supracellular level.
A group of cells situated at the front of the supracellular unit generally becomes the
leader cells in response to external cues and extend stable lamellipodia or filopodia
towards the substrate, whereas the follower cells situated at the rear extend
small transient cryptic lamellipodia. The stable protrusions possibly together with
the transient ones promote the formation of focal adhesions with the ECM to exert
traction forces towards the substrate. Application of these forces physically deforms
the matrix, creating a path for the entire cohort211,229. Leader cells can also secrete
matrix metalloproteinases that remodel the surrounding ECM, paving the way for
collective migration during cancer invasion. In addition to this traction-​based collective
migration, cells can adopt a propulsion-​based motility analogous to the amoeboid
mode detected, for instance, in colon cancer cell clusters231.
Complicating any classification scheme of cell migration is the fact that cells can
switch between modes of single-​cell migration and between single-​cell and collective
migration, depending on a variety of factors, such as tissue topology, ECM composition
and the degree of adhesion to it, as well as the presence of biochemical cues232,233.
For example, physical confinement and low adhesion enable slow-​moving fibroblasts
and epithelial cells to transition into a faster migration mode, where large stable blebs
are favoured by high cell contractility232. Similar stable bleb-​based fast migration is
observed in zebrafish progenitor cells induced by spatial confinement in vitro233 and
in vivo at transplantation-​induced wound sites of the embryo, where the cortical
contractility is elevated. Confinement-​associated plasticity in the migration mode is
also detected in leukocytes. For example, neutrophils with a genetically disrupted
branched actin network switch from multiple finger-​like protrusions to smooth
bleb-​based leading-​edge protrusions when exposed to confined microenvironments234.
Cancer cells can also adopt a rounded, bleb-​based migration mode in low-​adhesion 3D
environments or when their matrix metalloproteinase activities are disrupted. In addition,
physicochemical parameters such as hypoxia, which is a prominent feature of solid
tumours, have been shown to promote the transition of collectively invading cancer
cells into individually moving amoeboid cells, and enhanced cancer dissemination235.
The extreme plasticity in migration modes displayed by cancer cells may allow them
to adapt to many tissue environments and contribute to disease progression.
also known as electrotaxis) (Table 1). This framework is
Lamellipodia
Broad, sheet-​like protrusions deliberately generic to facilitate comparisons and con-
that contain branched trasts between different types of migration-​inducing cues
and linear actin filaments. and migration modalities. By comparing different forms
A variety of cell types, including of directed migration, we highlight the progress made in
fibroblasts, neural crest cells and
macrophages, use lamellipodia
the field and reveal gaps in our understanding of mole­
to explore longer distances cular underpinnings driving the directionality of cell
through the extracellular matrix. migration. This Review cannot comprehensively cover
530 | August 2021 | volume 22
0123456789();:
all aspects of this large topic. Thus, we will avoid descrip-
tions of new technologies for generating signal gradients
or for quantifying migration or mathematical/theoretical
models of this process, although we point to a few appro-
priate reviews of these topics (see refs9,10). Instead, we
will focus on how new findings provide insight into the
underlying principles of directed migration and suggest
questions to be addressed by future studies.
Four pillars of directed migration
To organize the large amount of information necessary
to understand directed migration in response to vari-
ous cues, we developed a generic, conceptual frame-
work of four events that must occur during all forms
of directed mig­ration, which we term the ‘four pillars of
directed mig­ration’ (Table 1). The pillars include genera­
ting the signal, sensing the signal, transmitting the signal
and executing the signal. For each pillar, we separate the
various forms of directed migration by cue except for
the fourth pillar, where we consider how the various sig-
nalling mechanisms converge on a common set of cell
translocation machineries.
Generating the signal
For directed migration to occur, a signal must first be
generated. This signal may be a transient cue such as
a diffusible chemical signal secreted into the environ-
ment meant to direct cells for a short burst of migra-
tion. Alternatively, the signal may involve a long-​lasting
change to the environment that guides cells for an
extended time, such as the generation of physical paths.
Chemotaxis. Chemotaxis is mediated by the generation
of diffusible cues. When these cues are presented uni-
formly, cells undergo chemokinesis, where they migrate
randomly with either higher speed and/or higher turn-
ing frequency relative to unstimulated cells11. However, if
the promigratory signal is presented in the form of a gra-
dient, directed migration occurs12 (Fig. 1a). The diffusible
agents that induce directed migration include a large and
diverse group of chemoattractants produced by different
sources. These comprise formylated peptides13, products
of the complement cascade14, phospholipid metabolites15
and a large family of chemokines and growth factors
that are derived from endothelial, epithelial and stromal
cells16. In addition, ATP and hydrogen peroxide have
been reported to act as autocrine signals to amplify
chemotactic signals 17–19. Furthermore, specialized
secreted proteins, such as Slits, netrins, semaphorins
and ephrins, are well-​known axon guidance cues20–23.
The diverse biochemical nature of these chemotactic
cues, with distinct diffusion coefficients and affinities for
their cognate receptors, presents considerable challenges
for the generation and maintenance of stable gradients
during chemotaxis.
A gradient can be established by simple diffusion from
a source or by regulating the removal of the attractant24.
Examples of mechanisms cells use to regulate gradient
formation and avoid receptor saturation (a situation
when cells are no longer able to perceive concentration
differences of the chemoattractant) include degrada-
tion of chemoattractants by enzymatic or proteolytic
www.nature.com/nrm
 Reviews

Filopodia
Finger-​like protrusions that
contain bundles of linear
F-​actin. Filopodia serve to
probe local environmental
cues, provide directionality
and maintain persistence
of migrating cells.
Stress fibres
Contractile arrays of actin
and non-​muscle myosin II
that are mechanically coupled
to the substrate through
integrin-​based focal adhesions.
breakdown, and endocytosis of cell surface-​b ound
chemoattractants via scavenger/decoy receptors that
specifically remove their ligand without initiating cell
polarity/migration signalling (reviewed in25) (Fig. 1b). For
example, it was established that a negative-​feedback loop
between CXCL12 and its receptor CXCR7 is required to
maintain optimal CXCL12 concentration in the zebrafish
posterior lateral line primordium26. Using a clever synthetic
approach, where GFP is used to generate diffusible gra-
dients, it was recently shown that combining the expres-
sion of non-​signalling decoy receptors with receptors
engineered to respond to GFP allows the synthetic GFP
gradient to generate normal growth and patterning of
the Drosophila melanogaster wing pouch27. In addition,
self-​generating gradients have recently been proposed
as an alternative mechanism to generate chemical gradi-
ents. In this case, migrating cells would secrete enzymes
that break down chemoattractants initially distributed
uniformly — as observed for Dictyostelium discoideum
cells migrating towards folic acid28, and melanoma and
pancreatic cells responding to lysophosphatidic acid29,30.
Such a mechanism can theoretically give rise to steep
gradients that work over long distances and convoluted
migratory pathways. Accordingly, with use of artificial
complex environments and mathematical modelling, it
was recently shown that the breakdown of attractants
allows D. discoideum and pancreatic metastatic cell
lines to navigate long, complex paths in a manner that

Table 1 | The four pillars of directed cell migration

Migration Cue Signal generation Signal sensing Signal transmission Signal execution
mode
Chemotaxis Diffusible chemical Simple diffusion GPCRs Classical signalling Leading-​edge
released from pathways involving small protrusions (all types)
Regulated removal RTKs
cells or deposited and large G proteins
by degradation of the Localized regulation
extracellular vesicles Other receptors, such as
chemoattractant or decoy PI3K of non-​muscle myosin
axon guidance receptors
receptors II and contractility
TORC2
Release of extracellular
PLA2
vesicles
MAPK/ERK
Haptotaxis Substrate-​bound ECM secretion and For ECM, integrins, Classical integrin signalling Biased protrusion
chemical cues such deposition but different adhesion pathways: Rho-​family generation through
as an immobilized structure impacts GTPases, FAK–Src, etc. a positive-​feedback
Binding of soluble factors
chemokine or ECM signalling outcome loop. Requires the
to a substrate (mostly Bound chemokine:
ECM) For substrate-​bound probably same as diffusible Arp2/3 complex
Exposing new sites on chemokines, regular cue
the substrate through receptors, but signalling
enzymatic action kinetics may be different
for different receptor–
ligand pairs
Durotaxis Differential substrate Passive: creating a stiff Integrins Unclear, but two Similar to other
compliance substrate by crosslinking mechanisms have forms of migration
Membrane tension
of ECM components or been proposed: role of but biased relative
and/or invagination
ECM deposition pure mechanics using to stiffness gradient
Focal adhesion actomyosin system
Active: cells or tissues
components or the involvement of
exerting a force on the
substrate that is sensed Actomyosin filaments mechanically triggered
by other cells signalling events
LINC complex
Topotaxis Geometric Preformed tunnels Cells adhere and conform Cell and nuclear shape Topology/
properties of the created by other cells to the topology and/or change may affect geometry biases
migration substrate the geometry of the both signalling and the force-​generating
Trails created by
irrespective of migration substrate with cytoskeleton but the mechanisms of actin
proteolytic ECM
mechanical or the help of focal adhesion mechanisms remain polymerization
remodelling
chemical properties components unclear and actomyosin
Topological features contractility
created by non-​lytic ECM Membrane curvature-​
deformation sensing proteins

1D fibrils such as bundles Nucleus deformation

of collagen
Topology of natural tissue
elements
Galvanotaxis Electric fields Ionic differences Electrophoretic Clustering of membrane Similar to chemotaxis
generated by movement of charged proteins/lipids must but biased relative to
transepithelial barriers surface proteins and activate signalling, but charge
such as in the skin, lipids within the plane the mechanisms remain
disrupted by wounding of the membrane unclear
Arp2/3 complex, actin-​related protein 2/3 complex; ECM, extracellular matrix; FAK, focal adhesion kinase; GPCR, G protein-​coupled receptor; LINC complex,
linker of nucleoskeleton and cytoskeleton complex; PI3K, phosphoinositide 3-​kinase; PLA2, phospholipase A2; RTK, receptor tyrosine kinase; TORC2, target of
rapamycin complex 2.

NAture RevIews | MolECular CEll BioloGy volume 22 | August 2021 | 531

0123456789();:
Reviews

a Signal generation
Chemotaxis Haptotaxis Durotaxis

Chemoattractant Collagen Low stiffness High stiffness

fibre
LOX

ECM protein
Chemoattractant
Enzyme Crosslinked ECM fibres
Chemical Release of ECM-bound
concentration chemoattractant
Topotaxis Galvanotaxis

–––––
Tissue Electric current
Confined tunnel Nanostructures wounding

MMP +++ +++ –––––

Tracks of Tracks generated by Transepithelial Electric field
aligned fibres proteolytic remodelling electric potential generation

b Gradient generation and stabilization

Through enzymes and decoy receptors Through EV release

Enzyme
GPCR

Endosome
Multivesicular
body with EVs
Decoy
receptor
EV establishing EV establishing
heterotypic homotypic gradient
gradient

Substrate compliance
The mechanical resistance
provided by non-​rigid
substrates (for example,
collagen gels) to the contractile
forces exerted by cells as they
engage the substrate.
Complement
Complement proteins are
products of the complement
pathway generally activated
as part of the innate immune
response to infection. Some
complement proteins, such as
C5a, act as chemoattractants
that guide leukocytes to sites
of infection.
Fig. 1 | Generating the signal. a | The diverse ways by which cues for directional migration are generated. During chemotaxis,
soluble chemoattractants released from bacteria or cellular sources diffuse to form chemical gradients. During haptotaxis,
extracellular matrix (ECM) proteins and chemokines released from cellular sources are deposited onto the ECM and
generate gradients of immobilized chemical cues. In some cases, ECM-​bound chemokines are released from the matrix
by cellular proteolytic activities (scissors) and provide soluble cues for chemotaxis. During durotaxis, gradients of stiffness
can be generated by lysyl oxidase (LOX)-​mediated ECM crosslinking. During topotaxis, the geometry of the existing tissue
structures, aligned fibres or tracks generated by proteolytic remodelling (via matrix metalloproteinases (MMPs), scissors)
or deformation provides directional signals. During galvanotaxis, electric fields generated at wounding sites as a result
of the loss of transepithelial potential provide guidance cues for cells involved in damage repair. b | Cartoon explaining
how stable gradients are generated and maintained during chemotaxis. Uniformly present soluble chemicals are either
degraded by enzymes or scavenged (via endocytic internalization) by decoy receptors to establish a gradient (left).
Cells release extracellular vesicles (EVs), such as exosomes carrying either identical (homotypic gradient) or distinct
(heterotypic gradient) chemical cues, to generate a stable secondary gradient (right). GPCR, G protein-​coupled receptor.
is dependent on attractant diffusibility, cell speed and that harbour motifs that bind to extracellular matrix
path complexity31. (ECM) components and can be later released by
Gradients can be propagated by means other than cell-​mediated proteolysis of the ECM component33.
simple diffusion 32. For instance, morphogens have Similarly, it has been shown that neutrophils migrat-
long been known to be secreted in precursor forms ing in 3D collagen matrices activate discoidin domain

532 | August 2021 | volume 22 www.nature.com/nrm

0123456789();:

Posterior lateral line
primordium
A group of cells that migrate
together from the ear to the tip
of the tail of zebrafish as they
periodically deposit primary
neuromasts.
Morphogens
Signal molecules that originate
from a tissue and diffuse to
generate a concentration
gradient. Morphogens exert
long-​range signalling effects
important for growth and tissue
patterning during development.
Advection fields
Fluid flows such as interstitial
flow in tissues that can create
an advection field or directional
transfer of molecules in the
liquid phase around cells,
which in turn can create
asymmetries in secreted
autocrine chemoattractants,
leading to autologous
chemotaxis. Advection fields
can also form in the cytoplasm.
Extracellular vesicles
A group of heterogeneous
vesicles (several nanometres to
micrometres in size) that carry
a variety of cargos, including
proteins, lipids and nucleic
acids, and are secreted by
cells to the extracellular
space to facilitate cell–cell
communication.
Exosomes
The smallest subtype of
extracellular vesicles, with
a size ranging from 50 to
150 nm. Exosomes are
generated as intraluminal
vesicles which are secreted to
the extracellular space when
intraluminal vesicle-​carrying
multivesicular bodies fuse with
the plasma membrane.
Caveolin
Integral membrane protein
family required for flask-​
shaped (caveola) membrane
structure formation. Caveolins
are also involved in membrane
trafficking, exocytosis,
endocytosis, extracellular
vesicle formation and
extracellular vesicle
cargo selection.
Rho-​family GTPases
A family of small proteins that
bind GDP or GTP and regulate
a wide array of downstream
signalling events. CDC42, Rac,
and RhoA are widely studied
members of this family
of proteins.
NAture RevIews | MolECular CEll BioloGy
receptor 2 (DDR2), which binds collagen I, and induces
the secretion of matrix metalloproteinases and the
release of collagen-​derived chemotactic peptides that
act in an autocrine manner to stabilize neutrophil
directionality34. The concept, referred to as ‘autologous
chemotaxis’, has also been reported to be involved dur-
ing the CCR7-​driven directed migration of tumour cells.
In this case, chemoattractant gradients were generated
from autocrine signals as a result of interstitial flow that
produced advection fields35,36.
During development, the transportation of signal-
ling molecules along filopodium-​like protrusions called
‘cytonemes‘ or ‘tunnelling nanotubes’ (thin cellular
protrusions involved in cell–cell communication) or
through transcellular transport (transcytosis) has been
reported to be involved in the generation of mitogen
gradients37,38. There is also evidence that the packaging
of chemoattractants in extracellular vesicles (in particu-
lar exosomes) importantly contributes to the genera-
tion of gradients during chemotaxis. In D. discoideum
cells, extracellular vesicles have been shown to contain
the machinery to synthesize and release the chemo­
attractant cAMP; these extracellular vesicles mediate the
relay of chemotactic signals during chemotaxis and
the alignment of cells in a head-​to-​tail fashion in a process
referred to as ‘streaming’39,40. In macrophages and den-
dritic cells, the secondary chemoattractant leukotriene
B4 (LTB4) has been reported to be present in exosomes
and promote migration41. In neutrophils, LTB4 is released
in response to primary chemoattractant stimulation and
acts in an autocrine and paracrine fashion to stabilize
neutrophil cell polarization and to relay signals to dis-
tant neutrophils42,43. Similarly, dendritic cell migration
was recently reported to depend on exosomes released
from lymphatic endothelial cells in a CX3CL1 (also
known as fractalkine)-​dependent fashion44. Moreover,
chemokine-​containing exosomes isolated from stressed
tumour cells have been reported to activate and induce
the migration of T cells45, and neutrophils have been
shown to package and release CXCL12 via secretory
vesicles, leaving behind CXCL12-​containing trails that
attract T cells to infection sites46. In these contexts, the
packaging of chemotactic cues in extracellular vesicles/
exosomes is poised to protect attractants from harsh
extracellular environments, degradation and/or rapid
diffusion. Furthermore, we envision that as the vesicles
are deposited, they can persist after the cells have left
the area and continue to deliver chemotactic cues to
generate long-​lasting secondary gradients to recruit dis-
tant cells to sites required for their action (such as sites
of inflammation for leukocytes) (Fig. 1b). In addition,
extracellular vesicles have also been reported to mediate
directional migration by regulating cell–ECM adhesion
assembly in tumour cells. In this case, autocrine secre-
tion of fibronectin-​coated exosomes at the leading edge
of cells expressing fibronectin receptors allowed them to
establish connections to the ECM, which became coated
with these extracellular vesicles47,48.
Haptotaxis. Haptotaxis is the sensing of surface-​bound
chemical cues (Fig. 1a). A primary haptotactic cue is
provided by the components of the ECM. A variety
0123456789();:
Reviews
of cells can secrete ECM proteins, such as fibronectin,
laminin and various collagens, into the environment to
form insoluble arrays that become migration substrates
or bind to existing substrates, thereby functioning as
migration cues for the cells themselves or for other cells
in the vicinity49,50. Unlike diffusible chemotactic cues,
ECM haptotactic cues tend to be relatively stable and
long-​lasting. As many components of the ECM can bind
to each other, their deposition can be iterative to create
complex mixtures of haptotactic cues51. In addition, cells
can locally degrade ECM components to further sculpt
remarkably complex migration environments52.
The specific molecular mechanisms of ECM secre-
tion remain poorly understood. Recent work suggests
that caveolin-​dependent regulation of exosome biogen-
esis is a key step in ECM secretion and deposition in
fibroblasts53, but the universality of this mechanism will
need to be explored in other cell types. Furthermore, ini-
tial cell engagement with the ECM can trigger the depo-
sition of fibronectin at the leading edge of cells through
a mechanism involving one of the Rho-​family GTPases,
CDC42 (ref.54). This creates a positive-​feedback loop for
cells to essentially lay down tracks and facilitate persis-
tent migration on ECM compositional gradients. Cell
engagement with the ECM can also modify the structure
of the array, whereby integrin-​dependent cell contacts
induce the reorganization of fibronectin fibrils55. An
interesting pathophysiological example of ECM depo-
sition occurs in the retina of patients with diabetes or
macular degeneration, where inappropriate deposition
of fibronectin leads to thickening of Bruch’s membrane
(the innermost layer of the retina) and inappropri-
ate neovascularization56. Such increased deposition of
ECM proteins could generate abnormal haptotactic cues.
However, increased ECM deposition will also change
the mechanical landscape of the microenvironment,
which could trigger a pathological durotactic response
as described later.
In addition to ECM haptotactic cues, cells secrete
factors such as chemokines or other guidance factors
that bind tightly to existing ECM arrays57, thus gen-
erating haptotactic cues that are sensed by direct cell
engagement rather than acting at a distance through
diffusion like during chemotaxis. Generation of these
cues is regulated by mechanisms similar to the ones
responsible for the generation of diffusible, chemotactic
cues. Nevertheless, as noted already, such ECM-​bound
chemokines can also be released to switch a haptotactic
cue into a locally functioning chemotactic cue58,59.
Durotaxis. In addition to sensing molecular cues, cells
have the ability to sense differences in substrate stiffness
and respond by migrating towards or away from areas
of higher stiffness (Fig. 1a). Such stiffness gradients have
recently been demonstrated in vivo in the embryonic
mouse limb bud60. The generation of these durotactic
cues requires changes in the mechanical environments
that cells encounter, which can persist for long periods
and influence the migration of cells for days or much
longer. For example, increased amounts of ECM deposi-
tion can often lead to changes in the mechanical proper-
ties of the local microenvironment, which can produce a
volume 22 | August 2021 | 533
Reviews
G protein-​coupled receptors
(GPCRs). A family of plasma
membrane receptors composed
of seven transmembrane
domains that couple to
heterotrimeric G proteins to
regulate responses mediated
by a variety of external signals.
Axon growth cone
Motile structure at the tip
of growing axons that guides
directed extension of the axon
and is important for patterning
of the nervous system.
534 | August 2021 | volume 22
hybrid haptotactic–durotactic–topotactic cue61. Another
means by which mechanical cues can be generated is
through mechanical modification of the existing ECM,
either stiffening it or relaxing it. For example, the lysyl
oxidase enzymes (LOXL1–LOLX4) can crosslink col-
lagen fibrils and other ECM components to render a
stiffer network62,63. Interestingly, this group of enzymes
is frequently misregulated in cancer and other disease
states such as during fibrosis, speaking to the importance
of mechanical control of the cell’s environment62. These
mechanical changes can influence the proliferation and
migration of tumour cells64 as well as surrounding stro-
mal cells such as endothelial cells in the vasculature65.
Conversely, matrix-​degrading enzymes such as matrix
metalloproteinases can relax or soften the environment
to further sculpt the mechanical landscape encountered
by cells66,67.
Topotaxis. Topotaxis is driven by biophysical cues,
where cells sense the topographical features of the sur-
rounding microenvironment. Natural tissue elements,
including aligned collagen fibres, muscle strands, nerve
fibres, vascular tracks and pores or tunnel-​like confined
trails within the ECM, often provide an anisotropic sur-
face architecture at nanometre or micrometre scale68,69
(Fig. 1a). Migrating cells tend to adapt their shape to
the available geometry of the surrounding substrate
to migrate in a preferred direction. However, it is impor-
tant to consider the cell types (taking into account their
unique properties) as well as the size of the confining
space when one is reflecting on how topological cues pre-
scribe directional choices to migrating cells. For instance,
it has been shown that migrating leukocytes, which
exhibit amoeboid movement, follow pre-​existing trails
in 3D reconstituted collagen matrices70,71. Unlike tumour
cells and fibroblasts, these leukocytes do not actively
break down the ECM. Instead, leukocytes migrating in
interstitial tissue undergo a robust shape change guided
by the matrix that induces transient deformation of the
collagen network, and squeeze through the preformed
trails of larger pore size and least resistance. By contrast,
cancer cells during tissue invasion frequently depend on
proteolytic remodelling of the ECM to create their own
trails, especially when encountering environments with
limited space and more resistance 69,72. However, non-​
proteolytic strategies, including ECM deformation, have
been reported in cancer cells with amoeboid features as
they make their way through tissue73. Depending on the
tumour type, a customized migratory approach may
also exist, where tumour invasion relies on protease-​
dependent tunnel formation in the matrix, which guides
the migration of leading tumour or non-​tumour stromal
cells (including fibroblasts, endothelial cells and smooth
muscle cells), whereas the follower cells are simply car-
ried along those tunnels without the need for active ECM
remodelling74. Further, it has been reported that cancer
cells during invasion can orient themselves parallel to
different topological features of the surrounding tissue,
and migrate directionally without requiring major ECM
remodelling, suggesting that cells are capable of sensing
complex topological features of the microenvironment
and tune their migratory response accordingly75.
0123456789();:
Galvanotaxis. The existence of electric fields around
cutaneous wound sites has been known since the
1840s76, and these fields can serve as cell migration cues
in a process called ‘electrotaxis’ or ‘galvanotaxis’ (Fig. 1a).
During the wounding process, the electric poten-
tial maintained by transepithelial resistance is short-​
circuited, and the resulting electric field can reach up
to 10 μA cm−2, a value in the range that can be sensed by
cells77. In addition to wounds, electric fields have been
documented during embryogenesis through transep-
ithelial ion transport78. These transient electric fields
are sensed by cells in their vicinity, often provoking a
directed migration response.
Sensing the signal
Once a signal has been generated, cells must be able to
sense the signal. In the case of chemotaxis, this is a fairly
straightforward process of receptor–ligand interactions.
However, the sensing step for other cues, such a mecha­
nical compliance of the substrate, is less straightforward
and involves more complex mechanotransduction path-
ways engaging both surface proteins and mechanically
coupled intracellular proteins.
Chemotaxis. The mechanisms underlying how cells
sense extracellular cues are, by far, best understood for
chemical cues in the context of chemotaxis (Fig. 2a).
Work using D. discoideum in the late 1980s first identi-
fied G protein-​coupled receptors (GPCRs) as the surface
receptor responsible for sensing cAMP — the main chemo­
attractant for D. discoideum chemotaxis79. This work
was followed by a flurry of reports in the early 1990s
showing that chemokines are also sensed by GPCRs80–83.
More than 50 distinct chemokines have been identified
in humans and are grouped into the CL, CCL, CXCL
or CX3CL subfamily, depending on the sequential posi-
tioning of highly conserved cysteine residues84,85. These
are recognized by ~20 known conventional chemokine
receptors, referred to as ‘CCRs’ or ‘CXCRs’, that share
25–80% sequence identity and exhibit the ability to
bind multiple chemokines within a given chemokine
subfamily83,86,87. In addition, some chemokines can bind
to atypical chemokine receptors, which are structurally
related to conventional chemokine receptors but do
not couple to signalling modules. Finally, formylated
peptides88, products of the complement cascade16, phos-
pholipid metabolites89 and the small molecules ATP and
ADP17,90 are all known to bind to GPCRs to mediate
their chemotactic activities, making GPCRs the main
molecular chemotactic sensors.
Receptor tyrosine kinases (RTKs) that interact with
growth factors, such as EGF and PDGF, are a second
class of receptors that mediate chemotaxis91. Whereas
GPCR-​mediated chemotactic signalling prevails in the
context of amoeboid chemotaxis, mesenchymal cells
often use RTK-​mediated signalling during directional
migration, perhaps reflecting fundamental differences
in physiological and environmental conditions encoun-
tered by each cell type92,93. In addition, axon growth cone
guidance, which is required for patterning the nervous
system, is mediated by several families of transmem-
brane receptors that bind to secreted guidance cues
www.nature.com/nrm
 Reviews

a Signal sensing
Chemotaxis Haptotaxis Durotaxis

Chemoattractant
receptor Chemoattractant
Chemoattractant receptor Chemoattractant LINC
receptor complex
Integrins
Immune Fibroblast Mechanosensing
cell components

Microtubules ECM protein

Collagen Crosslinked ECM fibres
Neuronal fibre
growth cone
Chemoattractant
receptor
Topotaxis Galvanotaxis
Protrusion–ECM
fibre alignment Electric current
Curvature
sensing BAR-family
Focal protein
adhesion
Tracks of aligned fibres Nanostructures +++ –––––
Shape Nuclear
adaptation deformation
Receptor clustering
via electromigration
Signal
Confined tunnel transduction

b Detailed machinery of durotaxis signal sensing

LINC
Membrane invagination complex Actin filament
during endocytosis Myosin
Mechanosensing
components Calcium ion
Stretch-activated
ion channel (Piezo)
Focal adhesion complex

Crosslinked ECM fibres

Fig. 2 | Sensing the signal. a | Various ways cells sense directional cues. During chemotaxis, cells sense the signal through
surface receptors (G protein-​coupled receptor (GPCRs), receptor tyrosine kinases or other transmembrane receptors),
which bind the soluble chemical cues. During haptotaxis, cells detect surface-​bound cues through integrin receptors and
GPCRs. During durotaxis, substrate stiffness is sensed by an array of mechanically coupled components located on the cell
surface, in the cytosol or at the nuclear envelope. During topotaxis, cells detect the geometry of available space and adapt
their shape by changing the orientation of membrane protrusions in parallel to the aligned extracellular matrix (ECM)
fibres, sensing topology-​induced membrane curvature by BAR-​family proteins, or gauging nuclear deformation resulting
from compression and shape change. During galvanotaxis, the electric field is sensed by electromigration of membrane
components (including signalling receptors) towards the cathode (+++) or the anode (− − − − −). b | Molecular machinery
for durotactic sensing. Cells sense gradients of stiffness using mechanosensors at the cell surface (including integrin
receptors in focal adhesions, invaginated membranes and stress-​activated ion channels), inside the cytoplasm (including
components of focal adhesions, actin filaments, microtubules and other mechanosensitive proteins) or at the nucleus
(LINC complex).

(such as Robo–Slit proteins or netrin–netrin receptors)
and mediate either attractive or repulsive responses94–96.
Neuronal guidance cues are also known to regulate
immune responses97, where, again, they can inhibit or
promote migration98–100.
Haptotaxis. Haptotactic cues are also sensed by cognate
cell surface receptors (Fig. 2a). Integrins are used to sense
haptotactic gradients composed of ECM components,
where the spatial organization or clustering of integrins
is key to their signalling properties101. Numerous studies

NAture RevIews | MolECular CEll BioloGy volume 22 | August 2021 | 535

0123456789();:
Reviews
Cell–cell junctions
Stable or dynamic sites where
borders of two neighbouring
cells contact each other.
Cell–cell adhesion receptors
and recruited adaptor proteins
are mechanically coupled to
the actin cytoskeleton.
Focal adhesions
Multiprotein assemblies that
physically connect extracellular
matrix components to the
intracellular actin cytoskeleton
through integrin clusters.
Integrin-​mediated adhesion
to extracellular matrix ligands
recruits a plethora of signalling
(Src and FAK) and structural
(talin, paxillin and vinculin)
molecules to focal adhesions.
Large, mechanically engaged
focal adhesions play a crucial
role in sensing mechanical
cues, while smaller, nascent
adhesions are critical for
sensing haptotactic cues.
Traction force
The stress vector at the
interface between a migrating
cell and its substrate.
LIM domains
Protein structural domains
named after the proteins
LIN-11, ISL1 and MEC-3.
A subset of these domains, such
as those found in the proteins
zyxin, paxillin and testin, bind
actin filaments in a mechanical
stress-​dependent manner.
LINC complex
Linker of nucleoskeleton and
cytoskeleton (LINC) complex is
a complex of nuclear envelope
proteins that connects the
cytoskeleton to the nuclear
lamina and is thus involved
in transferring signals from
sensing mechanical cues at
the cell surface or in the
cytosol into nucleus.
536 | August 2021 | volume 22
have shown that small, nascent adhesions at the leading
edge of migrating cells recruit a different subset of sig-
nalling and mechanical effectors compared with larger,
maturer adhesions further back under the cell body102.
Moreover, these small, nascent adhesions are critical for
ECM haptotaxis through a specific signalling pathway
described in the next section. In the case of surface-​
bound chemokines such as CCL21, the same GPCR
(CCR7) that senses the diffusible version of this cue is
used to sense immobilized gradients103. Interestingly,
dendritic cells following surface-​immobilized CCL21
gradients require non-​linear, exponential gradients for
haptotaxis and do not undergo haptotaxis on linear sur-
face gradients, suggesting differences in how the same
cue is sensed when it is diffusible or surface-​bound.
In theory, cells could use other receptors such as cad-
herins at cell–cell junctions to perceive haptotactic cues
presented by other cells, but this has yet to be reported.
Durotaxis. The mechanism of durotactic sensing is a
matter of intense recent interest (reviewed in104–106).
Unlike the cell-​impermeant chemical cues driving
chemotaxis and haptotaxis, which must be sensed by
cell surface receptors, mechanical force is not limited
by membranes. Thus, the ‘receptors’ or ‘sensors’ for duro­
taxis could theoretically be on either side of the plasma
membrane or even much deeper in the cytoplasm of the
cell, as long as these components are mechanically cou-
pled to the substrate. We envision that durotaxis results
from the ensemble activation of multiple mechanically
sensitive ‘receptors’ that act in concert to drive directed
cell migration.
Several candidate durotactic ‘receptors’ have been
identified (Fig. 2b) . Working from outside the cell
inward, the first candidate receptor are integrins107,108.
Several recent biophysical studies have demonstrated
that these surface proteins are sensitive to mecha­
nical load and are concentrated in structures relevant
for cell migration, such as filopodia, lamellipodia and
focal adhesions109,110. However, as some cells can migrate
by integrin-​independent mechanisms111, if they are
sensing substrate compliance, they may be using an
integrin-​independent mechanism. Another potential
source of mechanical sensing is the membrane itself,
which seems to prominently involve invaginations of
the membrane formed during either clathrin-​based
or caveolin-​based endocytosis. Indeed, several studies
have shown that these endocytosis pathways display
different dynamics and internalization frequencies
when cells are plated on uniform substrates of varying
compliance112–114. However, differences in endocytic
structures across single cells plated on stiffness gradi-
ents have not been shown. In addition, stretch-​activated
ion channels, such as PIEZO1/2, have been reported to
sense substrate stiffness115. One possibility is that as cells
apply traction force to the substrate on stiffness gradients,
these channels will become differentially activated and
produce local differences in migration-​relevant signal-
ling intermediates such as intracellular calcium. More
work will be required to test these ideas.
Moving just inside the plasma membrane, many
cytoplasmic components of integrin-​containing focal
0123456789();:
adhesions have been shown to be mechanically sensi-
tive, including talin116, vinculin117 and p130Cas118. When
focal adhesions are under differential mechanical load,
many of these proteins show altered conformation
and/or altered interactions with other cytoplasmic com-
ponents. However, many cells lack the large, stable focal
adhesion structures of mesenchymal cells, where these
phenomena have been mostly studied, and still display
sensitivity to substrate rigidity119. This suggests that
focal adhesions may not be a universal durotactic sens-
ing structure across all cell types. Regardless of whether
integrins are clustered in classic focal adhesion struc-
tures, they are still mechanically coupled to actin fila-
ments, and several recent studies have shown that actin
filaments themselves are sensitive to mechanical load by
differentially binding proteins containing LIM domains
depending on the mechanical conditions (varying
load)120,121. This differential response to mechanical load
could serve as a type of ‘durotactic receptor’, leading to
altered cytoskeletal dynamics and structure, as well as
alteration of signalling pathways. Recent work has also
shown that a subset of microtubules originating from the
Golgi apparatus are critical for durotaxis by regulating
focal adhesion dynamics122.
Finally, actin filaments and microtubules are con-
nected to the nucleus by the mechanically sensitive
LINC complex123,124, which could function as a sensor for
differential mechanical load on either side of the nucleus.
Indeed, differential positioning of the nucleus relative to
the rest of the cell has been linked to regulation of cell
polarization during scratch-​induced migration of epi-
thelial monolayers125 as well as cell locomotion through
piston-​like generation of pressure gradients in fibroblasts
migrating in a lamellipodium-​independent manner in
certain 3D environments126 (see the section Executing
the signal for details).
Topotaxis. We are only beginning to understand the
mechanisms for sensing topology (Fig. 2a). Similarly to
durotaxis, sensors of topological cues are mechanically
coupled elements that are located on either side of the
cell surface or within internal compartments, such as
the nucleus. Some of the sensing elements for topotaxis
(for instance, focal adhesions) may overlap with those
for both haptotaxis and durotaxis. Over the past dec-
ade, a large number of studies have monitored changes
in cell shape, protrusion shape, actin cytoskeleton and
motility in response to artificial matrix landscapes of
diverse topology. These landscapes are generally engi-
neered to house symmetric or asymmetric microstruc-
tures or nanostructures of the synthetic substrates that
are arrayed on a flat surface in specific patterns127–129
or 3D channels of open or closed ratchets to mimic
topological features of tissue architecture in vivo130.
However, studies aimed at teasing apart the mechanisms
of landscape sensing to the downstream intra­cellular
events, which translate into motility, are sparse. In a
few studies, cells were reported to sense topographical
features at nanometre to micrometre scales by extend-
ing leading-​edge protrusions. For instance, lamellipo-
dia in fibroblasts were shown to orient themselves
parallel to the micropatterned lines coated with ECM
www.nature.com/nrm

BAR-​family proteins
Bin/amphiphysin/Rvs161
domain (BAR) proteins are
membrane-​binding proteins
that aid in regulating
membrane shape.
Arp2/3 complex
A seven-​subunit protein
complex that possesses actin
nucleation and branching
activities leading to the
generation of branched
actin networks.
N-​WASP
Neuronal Wiskott–Aldrich
syndrome protein activates the
Arp2/3 complex and promotes
branched actin filament
formation.
Cortactin
A nucleation promoting factor
that activates the Arp2/3
complex and promotes
branched actin filament
formation.
NAture RevIews | MolECular CEll BioloGy
component, promoting elongated cell shape and direc-
tional migration131. In addition to parallel orientation, a
smaller size of the protrusions has been reported to pro-
mote directed migration, possibly by limiting migration
perpendicular to the direction of ECM patterns131. In
contrast to lamellipodia used by fibroblasts, neurons use
both transient, non-​aligned and stabler, aligned filopo-
dium populations to sense nanotopographical cues, and
to coordinate the signals that promote neurite outgrowth
along ECM-​coated lined patterns132. In addition to the
orientation of the structures, the availability of continu-
ous adhesive surfaces on the ECM-​coated aligned fibres
assists individual protrusions in sensing the topology133.
Such adhesion points give rise to the formation of
sequential focal adhesions along the continuous stretch
of fibres, which promote persistence of the protrusions
in a parallel orientation with respect to the aligned fibres.
Local deformation or curvature of the plasma mem-
brane at the interface of the cell and the substrate was
recently identified as the sensor of nanotopography134.
Topography-​induced membrane curvature serves as a
bridge between surface topography and intracellular
actin reorganization. Key pillars of the bridge include
curvature-​s ensitive BAR-​family proteins, particularly
FBP17, which recognize and accumulate within high-​
curvature areas present on either end of nanobars fab-
ricated to provide nanoscale topological cues. Localized
FBP17 then facilitates nucleation of F-​actin by acti-
vating key actin cytoskeleton modulators, including
the Arp2/3 complex, N-​WASP and cortactin134. The result-
ing branched F-​actin network accumulates at both ends
of the nanobars and undergoes rapid polymerization–
depolymerization cycles. Of note, curvature sensing by
FBP17 recruitment is restricted to a curvature diam-
eter of less than 400 nm, indicating that topography
sensing by curvature-​sensitive proteins is size specific.
Mechanistically, the identification of membrane curva-
ture as the topology sensor fills the gap in our under-
standing of how topological cues from the surface
translate into actin fibre reorganization inside the cells
and, hence, is groundbreaking.
Finally, a rather non-​conventional sensor of topo­
graphy is the cell nucleus. The nucleus is a large, bulky and
relatively rigid cellular organelle, repositioning of which
is rate limiting during migration in constrained envi-
ronments. Recent studies have revealed that topological
cues trigger changes in nuclear subcellular location and
shape, which have a significant impact on path finding
and cell migration71,135,136. In the case of immune cells
using amoeboid migration, cytoskeletal forces position
the nucleus in the front portion of the cells. This allows
cells to sample densely packed tissue, gauge the available
space and choose the path of least resistance71. Notably,
two recent studies proposed that the nucleus acts as
an internal ruler that interprets cell shape in confined
spaces, and facilitates rapid movement136,137. Specifically,
it was shown that when the level of confinement is
increased above a certain point sensed via nuclear
deformation, cells exert an active contractile force. This
contractile response was connected to the progressive
expansion and unfolding of the nuclear envelope with
increased confinement. A completely unfolded nuclear
0123456789();:
Reviews
envelope triggered a contractile response that allowed
cells to resist the physical compression and squeeze out
of the confined space. Such a specific adaptive response
tailored by the nucleus may be of particular importance
for immune cell patrolling through dense tissues, for
progenitor cells migrating through a densely packed
cell mass during embryonic development or even during
cancer cell invasion.
Galvanotaxis. How cells sense electric fields has been
debated for several decades138. Unlike mechanical cues,
which can be sensed either at the cell surface or inside
the cell, electric fields must be sensed outside the plasma
membrane due to its high electrical resistance. The two
predominant models for this sensing are membrane
depolarization and electromigration of surface proteins.
Recent reports favour electromigration of surface pro-
teins as a mechanism of galvanotactic sensing139,140. In
these studies, the charge on the extracellular domains of
model proteins, as well as some lipids and carbohydrates,
resulting from the application of an electric field induces
electrophoresis of these molecules within the plane
of the plasma membrane, either towards or away from
the cathode140,141 (Fig. 2a). Moreover, it has been shown
that galvanotaxis itself is sensitive to extracellular pH,
which likely changes the charge state of proteins through
protonation139.
Transmitting the signal
In the third pillar of directed migration, the signal must
be transmitted from the sensor to the machinery nec-
essary to move the cell. In some cases, this occurs via
polarized second messenger pathways, but in other
cases the signal transduction machinery and the motil-
ity machinery may overlap, such as during durotaxis.
During the transmission step, a weak signal (in the form
of a shallow gradient of the cue) is often amplified to
produce a robust cellular response.
Chemotaxis. For cells to migrate, their cytoskeletal
machinery must be polarized142. The important ques-
tions are how shallow gradients of extracellular chemi-
cal cues can be transformed into steep polarized cellular
responses, and at what point in the signalling cascade
are responses confined to a defined subcellular loca-
tion. The introduction of GFP technology and live-​cell
imaging have been critical in addressing this question.
It was established that chemotactic GPCRs remain uni-
formly distributed in D. discoideum cells and neutrophils
undergoing chemotaxis143,144, suggesting that the signals
leading to the segregation of the cytoskeletal machin-
ery are downstream of receptor occupancy. However, in
lymphocytes, chemokine receptors have been reported
to be clustered at the front of cells145. In mesenchymal
tumour cells undergoing chemotaxis, the EGF receptor,
like GPCRs, is also distributed homogeneously on the
plasma membrane, but accumulates in endocytic vesicles
on the side of the cell exposed to the high concentration
of the chemoattractant146. Because the EGF receptor sig-
nalling can continue from internalized endosomes, this
suggests that polarized receptor signalling could occur
for RTKs in mesenchymal chemotaxis147.
volume 22 | August 2021 | 537
Reviews

a Polarization of intracellular b Signal transmission Signal sensing by receptor

signalling molecules
Signal transmission
Shallow gradient of directional cues RhoA PI3K TORC2 MAPK PLCγ

ROCK activation PtdIns(3,4,5)P3 DAG enriched at front

Rear Front enriched at front

PH domain-containing
MLCK proteins PKCα activation
Steep gradient of polarized
intracellular signalling molecules Rho-family GTPases
Myosin II light chain Myosin II inhibition
phosphorylation Arp2/3 complex at front

• Actin polymerization • Actomyosin contractility

• Front protrusion • Asymmetric force generation

c Signalling pathways triggered

by haptotactic cues
Fibronectin
(ECM)
Haptotaxis Chemoattractant Integrin
receptor cluster
Integrins
P
WAVE, Kindlin FAK P
WASP NPFs Src P
Arp2/3
Actin
ECM protein P
Chemoattractant
Collagen
fibre Rac Rho GEFs

Fig. 3 | Transmitting the signal. a | Cartoon depicting how shallow gradients of extracellular directional cues are
transmitted into steep gradients of intracellular signalling molecules at the front and rear of cells. b | Flow chart describing
how various signalling pathways activated by sensing of directional cues lead to changes in the cytoskeletal machinery.
c | Cartoon highlighting the molecular machinery that transmits haptotactic signals. Sensing gradients of fibronectin
through integrin engagement activates non-​receptor tyrosine kinases: focal adhesion kinase (FAK) and Src-​family
kinases. These kinases phosphorylate a variety of substrates, triggering the formation of new protein complexes, including
guanine nucleotide exchange factors (GEFs), and the activation of Rac, which leads to branched actin network formation
through Arp2/3 complex activation via the nucleation-​promoting factors (NPFs) WASP and WAVE, ultimately generating
lamellipodial structures. DAG, diacylglycerol; PH, pleckstrin homology; PI3K, phosphoinositide 3-​kinase; PtdIns(3,4,5)P3,
phosphatidylinositol 3,4,5-​trisphosphate.

Pleckstrin homology (PH)
domain
Small protein domains of
approximately 120 amino
acids that are known to have
phosphoinositide-​binding
specificity.
DOCK–ELMO
A protein complex consisting
of an adaptor protein, ELMO,
and a Rac-​specific guanine
nucleotide exchange factor,
DOCK.
Nevertheless, from findings in amoeboid cells it is
clear that the polarization of signalling molecules at the
front and rear of cells undergoing chemotaxis occurs
downstream of sensing receptors and upstream of the
cytoskeletal machinery (Fig. 3a,b) (although the flow of
actin may also contribute to the polarization of signal-
ling molecules and hence cytoskeletal rearrangements
may also fine-​tune the distribution and thus the recep-
tion of the signal)148. Indeed, with use of probes that
specifically label lipids downstream of phosphoinos-
itide 3-​kinase (PI3K), it was shown that phosphatidy-
linositol 3,4,5-​trisphosphate lipids are spatially restricted
to the leading edge of D. discoideum, neutrophils and
fibroblasts undergoing chemotaxis149–151. These polar-
ized phosphatidylinositol 3,4,5-​t risphosphate sites,
which are dependent on Ras signalling (reviewed
in152), are then poised to spatially recruit a subset of
pleckstrin homology (PH) domain-​containing proteins
that regulate actin assembly through the regulators of
Rho-​family GTPases, such as Rho guanine nucleotide
exchange factors, and DOCK–ELMO 153–155. However,
pharmacological inhibition or genetic ablation of PI3K
does not completely inhibit chemotaxis, particularly in
steep chemotactic gradients152,156,157, and several parallel
pathways have been reported to regulate D. discoideum
and neutrophil chemotaxis, including TORC2 (refs158–161),
phospholipase A 2 (refs 162–166) and MAPK/ERK 167–170. In
fibroblasts undergoing chemotaxis towards the RTK
ligand PDGF, the phospholipase PLCγ appears to be
crucial for this process through localized hydrolysis of
phosphatidylinositol 4,5-​bisphosphate to yield diacyl­
glycerol at the leading edge facing the highest concentra-
tion of PDGF156. This stable enrichment of diacylglycerol
triggers the localized activation of the kinase PKCα
and the subsequent inactivation or inhibition of myosin II
through non-​canonical phosphorylation of Ser1/Ser2

538 | August 2021 | volume 22 www.nature.com/nrm

0123456789();:

TORC2
Target of rapamycin complex 2
is composed of seven conserved
subunits and is involved in
regulating proliferation,
survival, cell migration and
cytoskeletal reorganization.
Phospholipase A2
An enzyme that cleaves
phospholipids to give rise
to lipid products (arachidonic
acid or lysophosphatidic acid)
that either have the ability
to regulate signalling events
or are substrates in the
generation of bioactive lipids.
MAPK/ERK
A group of protein kinases
that transduce signals from
cell surface receptors to
the nucleus.
Förster resonance energy
transfer
A mechanism describing
energy transfer between two
light-​sensitive molecules.
NAture RevIews | MolECular CEll BioloGy
on the regulatory light chain, thereby creating asymme-
try in myosin II-​mediated contractility. Furthermore,
in carcinoma cells migrating towards the RTK ligand
EGF, amplification of the signal has been reported to
occur through the activation of the actin-​s evering
protein cofilin by PLCγ thereby allowing asymmetric
actin polymerization required for protrusion towards
the EGF gradient171. Finally, positive-​feedback and
negative-​feedback mechanisms centred at the regu-
lation of signal transduction via the Ras–PI3K–ERK
pathway have been reported to be involved in reg-
ulating the modes of migration in D. discoideum172
and the metastatic potential of epithelial cells173. Such
self-​organizing excitable signal transduction activities
were proposed to underlie the control of the extension
of actomyosin-​based protrusions and were suggested to
have a key role during development (reviewed in142).
Importantly, although many aspects of chemotactic
signalling are common in amoeboid and mesenchymal
cells, the fundamental difference in migration behav-
iours between these two types of cells, with specific
mechanisms, timescales and dynamics, requires distinct
signalling pathways that are not fully understood92,93.
Haptotaxis. The transmission of haptotactic signals
and the transmission of chemotactic signals are similar,
involving traditional Rho-​family GTPase signalling path-
ways. The most well-​studied form of haptotaxis is sensing
gradients of ECM proteins through integrin engagement
(Fig. 3c). In this case, integrin engagement leads to the
activation of the non-​receptor tyrosine kinases FAK
and Src-​family kinases174. These kinases phosphorylate
a variety of substrates, triggering the formation of new
protein complexes including those involving guanine
nucleotide exchange factors, such as β-​PIX and TIAM1,
and the activation of Rac, which leads to the formation of
lamellipodial structures. When these lamellipodia pro-
trude up the gradient of fibronectin and encounter more
ECM ligands for the integrins, a positive-​feedback loop is
established that is critical for haptotaxis of fibroblasts on
gradients of fibronectin. However, it is not clear whether
signal amplification akin to what happens during chemo­
taxis occurs or is required during ECM haptotactic
signalling. In the case of haptotactic migration on gra-
dients of surface-​bound chemokines, the presumption
is that the signalling pathways activated by the cognate
GPCRs are the same as the signalling pathways activated
by the diffusible version of the chemotactic cue, but this
presumption needs further experimental validation.
Interestingly, one study reported differential regulation
of CCR7 towards haptotactic ECM-​bound gradients of
CCL21 versus chemotactic, diffusible CCL21 (ref.103).
This suggests that a cell response to haptotactic versus
chemo­tactic cues, even when involving the same receptor/
pathway, may be specifically modulated to adjust cell
behaviour to the migratory context.
Durotaxis. Delineating how durotactic signals are
transmitted is challenging as the ‘receptor’ or ‘recep-
tors’ responding to durotactic cues are not definitively
known. Furthermore, the distinction between sensing
and transmitting a mechanical signal could overlap
0123456789();:
Reviews
considerably, and thus is not nearly as clear-​cut as dur-
ing other directed migration types, such as chemotaxis.
However, the signal transduction mechanisms involved
during durotaxis can be broadly grouped into two
classes, depending on the cell type and environmental
context.
First, mechanically sensitive enzymes and proteins
can generate traditional second messengers such as
intracellular calcium changes. The best studied examples
of these are the non-​selective PIEZO1/2 stretch-​activated
ion channels175. These proteins have been linked to a
wide variety of mechanosensation events but, to our
knowledge, have not yet been specifically tested during
durotaxis. Cytoplasmic enzyme kinetics can also be con-
trolled by mechanical load176, which could theoretically
lead to altered second messenger signalling, but this has
not been documented during mechanically triggered cell
migration.
Second, mechanical force can result in protein con-
formational changes, such as unfolding, or changes in
biophysical properties, such as catch-​bond behaviour,
where mechanical load increases bond strength. One of
the first proteins shown to display a mechanically sen-
sitive conformational change and altered binding part-
ners was p130Cas, an adaptor protein regulating tyrosine
kinase-​based signalling related to cell adhesion118. This
paradigm of molecular stretching under mechani-
cal load has strongly influenced how we think about
mechanosensing177. This has been further reinforced by
the generation of Förster resonance energy transfer-​based
molecular tension biosensors, including those of integrin
adhesion-​associated vinculin and talin, which enabled
the visualization of local changes in tension that may
constitute signals for directed migration117,178–181. Both
vinculin and talin show F-​actin binding that is increased
under mechanical load182,183. This catch-​bond binding for
both proteins is asymmetric, with a bias towards binding
the pointed (minus) end of the actin filament. In nascent
adhesion structures at the leading edge, the proximal
actin networks — which generally face the membrane
with their barbed (plus) ends — are flowing backwards
by retrograde flow from the leading edge. This polar-
ized flowing population of filaments can be engaged by
vinculin and talin associated with the nascent adhesions
due to their asymmetric catch-​bond properties, possibly
contributing to localized adhesion maturation and hence
directional migration. The next step will be to observe
how these local, tension-​dependent conformational
changes translate to more global mechanotransduction
networks across single cells migrating on gradients of
substrate stiffness.
Topotaxis. How guidance signals from tissue topogra-
phy are transmitted within cells is not clearly defined.
One can speculate that it will involve calcium, in light
of the well-​recognized role of calcium in converting
external information into biological signals that subse-
quently lead to actin polymerization. Enhanced intra-
cellular calcium activity has been detected in astrocytes
seeded onto a micropatterned surface184. Astrocytes were
found to be elongated and aligned along the direction of
the grooves accompanied by frequent calcium peaks in the
volume 22 | August 2021 | 539
Reviews
Pseudopodia
Protrusive structures in
amoeboid cells generated
by branched and linear actin
filament arrays in the leading
edge and aligned with the
direction of movement.
Ena/VASP
Enabled/vasodilator-​stimulated
phosphoproteins are actin
polymerases that drive actin
filament elongation and
antagonize filament capping,
leading to the generation
of linear actin filaments.
Formin
A group of actin polymerases
that drives the formation
of linear actin filaments.
540 | August 2021 | volume 22
aligned cells relative to randomly oriented, rounded
cells on flat surfaces184. Whether such calcium activity
also tunes cell motility on such patterned surfaces is less
clear. It has been partly addressed using surface topol-
ogy with different degrees of confinement that mimic
narrow channels and fibre-​like tracks of natural ECM
in vivo. In one of the recent studies, elevated intracellu-
lar calcium concentration was found to be important in
mobilizing cells through confined microchannels, where
the confinement-​driven force was transmitted inside the
cells as calcium influx via activation of PIEZO1 (ref.185).
However, cell migration in the study was induced in
the presence of a chemotactic source, indicating the
need to further investigate whether topology or con-
fined space-​driven calcium influx controls cell motility
independently of chemical cues.
Furthermore, PIEZO1 activation was recently shown
to be regulated by the geometric features, such as rough-
ness and stiffness, of the surrounding substrate when
force was applied externally by substrate deflection at
cell–substrate interfaces186. For instance, when indi-
vidual pillars of a deformable micropillar array were
deflected, the amplitude of the PIEZO1-​mediated cur-
rent was higher in cells surrounded by sparsely arrayed
pillars, which led to less substrate roughness than for
the densely arrayed pillars. Notably, the amplitude of the
PIEZO1-​mediated current induced by pillar deflection
decreased when cells contacted substrates with greater
stiffness but relatively low roughness186. Whether mod-
ulation of PIEZO1 channel activation by substrate
mechanics affects calcium spatio-​temporal dynamics
and ultimately cell migration in response to various
degrees of compressive, tensile and shear forces relevant
in physiological contexts, such as dense tissues or blood
capillaries, remains to be addressed.
There is also evidence that topotactic cues, and in
particular cell compaction, are sensed via the nucleus,
which will inevitably undergo deformation in this con-
text, and that both calcium and the lipid-​based second
messenger arachidonic acid are essential to transmit this
signal to the cytoskeleton71,136,137. In this mechanism,
nuclear envelope stretching and unfolding resulting from
the compressive force of compaction is associated with
calcium release from internal stores, possibly through
stretch-​sensitive calcium channels. This then triggers
the redistribution of the calcium-​dependent phospho-
lipase A2 (cPLA2) from the cytosol to the stretched
nuclear envelope. Subsequent activation of cPLA2 leads
to release of arachidonic acid from the nuclear envelope
phospholipids. cPLA2-​mediated release of arachidonic
acid is critical for the biosynthesis of lipid mediators,
including LTB4, which have well-​established roles in
activating myosin contractility and immune cell migra-
tion in an autocrine and paracrine manner42,187. Notably,
LTB4-​synthesizing enzymes also reside in the nucleus,
which thereby identifies the nucleus as a dynamic hub for
both sensing elements and second messenger production
during cell motility in response to confined topology.
Galvanotaxis. Cells in electric fields activate a wide
variety of intracellular signalling pathways, including
AKT, Src-​family kinases, MEK–ERK and JAK1 (ref.77).
0123456789();:
Consistent with this finding, galvanotaxis is sensitive
to many pharmacological inhibitors and genetic per­
turbation of intracellular signalling pathways 77,188.
In this way, galvanotaxis is most similar to chemot-
axis in terms of signal transmission. Consistent with
this idea, a large-​scale genetic screen for D. discoideum
mutants defective in galvanotaxis revealed an impres-
sive overlap of identified genes with those previously
implicated in chemotaxis188. The activation of a wide
array of signalling pathways may arise due to the
electromigration-​based clustering of a range of cell sur-
face receptors on either side of the cell (relative to the
electric field) that somehow activates downstream, intra-
cellular signalling pathways leading to polarized signal
transduction139,140. Indeed, recent data indicate that local-
ized, photo-​induced clustering of GPCRs can induce
ligand-​independent signalling189. Interestingly, in kerato­
cytes, inhibition of one of the kinases commonly asso-
ciated with galvanotaxis, PI3K, can actually reverse
the polarity of galvanotaxis, leading to migration of
cells towards the anode rather than the cathode139.
Although the mechanism of this reversal is not known,
it may indicate that electric field-​induced signalling is
being activated on both the anode-​facing side and the
cathode-​facing side of the cell, resulting in a ‘tug of war’
for cell polarization decisions that can be influenced by
perturbation of intracellular signalling.
Executing the signal
The translocation of cells, individually or in groups,
relative to their environment requires the generation of
asymmetric or polarized forces relative to the substrate.
There are several sources of this force generation, but a
key concept is that directed migration-​promoting sig-
nalling biases these forces either towards or away from
environmental cues. Directed cell migration cues do not
activate special mechanisms of cell translocation but bias
the migration and cell polarity machinery that operates
during normal, random cell migration12. Depending
on the mode of migration used by a particular cell type
(Box 1), multiple directed migration cues can converge
on a common set of force generation mechanisms lead-
ing to cell translocation. Thus, with this fourth pillar of
directed migration, we will not separate the directed
migration forms by the environmental cue; rather, we
will discuss the different forms of asymmetric force gen-
eration activated during directed migration (Fig. 4) and
describe how the signal transmission step regulates this
force generation.
Protrusion-​based mechanisms. One of the character-
istic features of most migrating cells is that they have
some kind of protrusive structure at their leading edge,
which is aligned with the direction of movement (Fig. 4a).
With a few notable exceptions, such as nematode sperm
cells190, most protrusions are driven by the polymeriza-
tion of actin filament arrays191. In amoeboid cells, these
are known as pseudopodia, and the actin is a mixture
of branched actin networks produced by the activation of
the Arp2/3 complex and linear actin arrays produced
by Ena/VASP and formin proteins192. In fibroblasts and
other more adherent cells, the protrusions often take on
www.nature.com/nrm
 Reviews

a Protrusion-based mechanisms Branched b Contractility-based mechanisms

actin network Stress fibre based
Contractile
Actin Actomyosin force
network forming Actomyosin
Pseudopod stress fibre bundle

Actomyosin Myosin Myosin

network Integrin
Focal adhesion
complex
Collagen fibre
Lamellipodium
Blebbing based
Actin cortex
Filopodium Myosin Polarized New membrane–
Linker Asymmetric blebbing cortex attachments
protein membrane
delamination
Local from the cortex
Linear increases
actin filament in cortical
tension
Local rupture
of the cortex

c Alternative mechanisms
Nuclear piston based Friction based Molecular paddling based
Low pressure Actin cortex Transmembrane
zone at the back protein anchored
of the nucleus to the crotex

Back Front

High-pressure
Confined 3D space zone in front Cortex
of the nucleus retrograde
Cortex flow
retrograde flow Advection

Fig. 4 | Executing the signal. Depiction of how asymmetric force is generated through protrusion-​based, contractility-​
based or alternative mechanisms. a | Formation of protrusions such as pseudopodia, lamellipodia and/or filopodia is driven
by branched and linearly polymerized actin networks. b | Contractility-​based mechanisms of migration (characteristic
of mesenchymal cells) depend on the establishment of stress fibres, where a contractile array of actomyosin networks is
mechanically coupled to the substrate through integrin-​based focal adhesions (top). Many cell types, in particular in the
in vivo context, migrate via extension of blebs (bottom), which are generated through local increases in cortical tension
on the non-​blebbing side of the cell (marked with arrows) and asymmetric membrane tearing (delamination), or by local
rupture of the cortex, or both. c | Cells moving through a confined space use the nuclei as a piston to generate zones
of high cytosolic pressure at the leading edge. Cells moving in the absence of substrate adhesion depend on friction
between cells and the environment, generated by retrograde flow of the cortical actin. Cells can also use molecular
paddling to swim through the environment using horizontal rearward flow of transmembrane proteins anchored to the
actin cortex (advection).

SCAR/WAVE
Suppressor of cAR/WASP
family verprolin-​homologous
protein is a nucleation-​
promoting factor that activates
actin nucleation activity of the
Arp2/3 complex.
a flatter appearance and fan-​like architecture (referred
to as ‘lamellipodia’) that is more dominated by Arp2/3-​
branched actin, although they also contain linear
actin microspikes that can extend beyond the edge as
filopodia193. Some cells, such as neuronal growth cones,
have protrusions that are almost entirely formed by
filopodia, containing bundled linear arrays of actin with
concentrated actin polymerases such as Ena/VASP and
formins at their tip194. Of note, these different protru-
sions are not finite entities and are highly plastic and can
dynamically interconvert over time, such as when filopo-
dia direct where lamellipodia can form195 or when lamel-
lipodia yield clusters of filopodia196. The proportion of
actin network types within protrusions (branched versus
linear) is probably more important than the historical
naming convention for the protrusion type.
Over the years, many lines of evidence have linked
the formation of protrusions with various directed
migration cues193,197. For example, one of the most sat-
isfying connections between signalling pathways and
protrusions is the activation of the Arp2/3 complex, via
Rho-​family GTPases and nucleation-​promoting factors
such as SCAR/WAVE, a topic that has been extensively
reviewed elsewhere191,198. However, several groups have
shown that cells lacking the Arp2/3 complex can still
undergo chemotaxis in various contexts, albeit with
reduced migration efficiency197,199–201. This indicates
that the Arp2/3-​branched actin pathway is not strictly
required for chemotaxis in all circumstances and points
towards other possible mechanisms of asymmetric force
generation.
Other mechanisms responsible for triggering or
tuning protrusions have been identified. In the case of
haptotaxis, the activation of the Arp2/3 complex and
the subsequent generation of branched actin networks
at nascent integrin adhesions requires both FAK–Src

NAture RevIews | MolECular CEll BioloGy volume 22 | August 2021 | 541

0123456789();:
Reviews
Calpain
Calcium-​activated cysteine
protease that cleaves adhesion
complex proteins.
Border cell
A specialized cell type
that migrates as a group
through the egg chambers in
Drosophila melanogaster.
Blebs
Spherical membrane
protrusions that rely on myosin-​
based contraction and pressure-​
driven cytosolic flow. Bleb-​like
protrusions are commonly
used for motility by amoebas
and embryonic cells. However,
leukocytes and tumour cells
can use blebbing motility
especially in 3D environments
under confined conditions.
Reynolds number
A dimensionless number
important in fluid mechanics.
Cellular scales are inherently
a low Reynolds number
environment where inertia and
momentum are negligible
and thus movement requires
strategies different from those
for human-​relevant length
scales.
542 | August 2021 | volume 22
and the RAC1 GTPase, integrated through the WAVE cluster of cells was shown to have a mechanically con-
regulatory complex174,197 (Fig. 3c). In the case of neuronal tinuous band of actomyosin contractility around the
growth cones responding to chemotactic guidance cues, periphery of the cluster that can be locally inhibited by
emerging evidence suggests that monoubiquitinylation chemotactic cues at the front to produce a form of ‘rear
of VASP mediated by the ubiquitin ligase TRIM9 helps wheel’ drive motility213.
tune the activity of this actin polymerase at the tips of At the single-​cell level, localized control of contrac-
filopodia202. In the case of durotaxis, both Arp2/3-​based tility (either positive or negative) has a profound effect
lamellipodia203 and filopodia204 are required to sense and on polarization and directed migration214. In fibroblasts
respond to varying substrate stiffness, although whether migrating in a gradient of the chemoattractant PDGF,
these protrusions are regulated through traditional, sec- local inhibition of myosin II at the leading edge is criti­cal
ond messenger-​type signalling pathways that are respon- for chemotaxis156. At the opposite end of the cell, recent
sive to substrate stiffness remains to be determined. One studies in neutrophils and keratocytes have shown that
consistent mechanism of protrusion control is the split- control of rear contractility governs the direction of
ting of existing protrusions and the selective stabilization cell movement215,216. Even in cells that lack strong cell–
of the protrusion that is oriented more strongly towards substrate or cell–cell adhesions, such as amoeboid
the gradient of the signal. This has been demonstrated tumour cells, local contractility may have a key role in
in the chemotactic migration of both D. discoideum the generation of cellular blebs through local increases
and fibroblasts205,206, and may reflect a common strat- in cortical tension on the non-​blebbing side of the
egy for many forms of directed migration. Finally, in cell, local relaxation of the cortex to permit blebbing
cells undergoing topotaxis, including D. discoideum, or both217. However, the signals upstream of polarized
neutrophils and breast cancer cells, it was observed that blebbing are incompletely understood218.
asymmetric microstructures of the substratum induce
actin waves, which propagate unidirectionally, defining Alternative mechanisms via pistons, ridges and molecu-
the direction of cell translocation. Curiously, the waves lar paddles. Beyond actin-​based protrusions and acto-
could propagate in different directions, depending on myosin contractility, cells have other ways to generate
the cell type, and this was associated with the establish- asymmetric traction force relative to their environment
ment of different focal adhesion patterns and differences and move (Fig. 4c). However, most of these have not
in membrane dynamics and protrusion extension, which been definitively linked to specific directed migration
were linked to distinct local cell cortical plasticity in cues. Cells moving through confined environments
different cell types129,207. can use their nucleus as a piston to generate cytoplas-
mic pressure gradients at the leading edge to facilitate
Contractility-​based mechanisms. The other main gen- migration126. Possible cellular regulation of this ‘nuclear
erator of asymmetric force relative to the substrate for piston’ mode could involve the association of the nucleus
migrating cells is contractile arrays of actin and non-​ with lateral membrane or localized osmotic control.
muscle myosin II (ref.208) (Fig. 4b). In the case of firmly Other mechanisms are observed in immune cells,
adherent cells, such as mesenchymal cells, these arrays which do not require integrins for their migration in 3D
of contractile actin form stress fibres that are mecha­ environments111. In one recent study it was shown that
nically coupled to the substrate through integrin-​based T cells can translocate using friction generated between
focal adhesions. Asymmetric strengthening or weak- the cell and its environment, which is coupled to cellu-
ening of these adhesion structures provides a dynamic lar translocation through retrograde flow of the actin
way to control the direction of cell migration. Indeed, cortex219. In another recent study, non-​adherent leuko-
many signalling pathways have been linked to adhe- cytes were shown to ‘swim’ through their environment
sion turnover, including phosphorylation of specific using a fascinating ‘molecular paddling’ mechanism,
adhesion proteins209 and the myosin II regulatory light which is mediated by transmembrane proteins coupled
chain156, selective proteolytic cleavage of adhesion pro- to the actin cortex and moving by advection across
teins by calpain210 and relaxation of adhesions through the cell surface in a coherent manner relative to their
contacts with microtubules68. Remodelling of focal environment220. This is a remarkably efficient migration
adhesions almost certainly underpins durotactic migra- strategy, with cell speeds similar to those of adhesion-​
tion in many cell types122,209 but may also be relevant for based migration. Such mechanisms could be of great
chemotactic migration of fibroblasts156. importance for migration through in vivo environments,
In the case of cells with robust cell–cell adhesions, given that cellular environments are characterized by a
such as sheets of epithelial cells, myosin-​based con- low Reynolds number221 — meaning that at cellular scales,
tractility can also be a potent mechanism to direct inertia and momentum are negligible and thus move-
migration211,212. In this case, the contractile arrays are ment requires strategies different from those for human-​
linked to the cell–cell adhesion sites and control the relevant length scales. However, it is unclear whether this
shape of cells relative to their neighbours, as well as coor- paddling mechanism is used only for random migration
dinate the transmission of mechanical force across mul- or is also responsive to external environmental cues.
tiple cells. Cells with strong intercellular junctions often
move collectively as groups, as in the case of border cell Conclusions and perspective
migration in D. melanogaster and neural crest migration in Directed migration has been studied for many decades,
vertebrates. In recent work on neural crest migration but our understanding of the mechanisms underpinning
in zebrafish and Xenopus laevis embryos, the migrating this process has increased dramatically only over the past
www.nature.com/nrm
0123456789();:
 Reviews

Box 2 | Key open questions for specific modes of directed cell migration
Chemotaxis
How are stable gradients of diffusible cues generated and maintained in tissues with
interstitial fluid flows as in most multicellular organisms?
Haptotaxis
Is haptotaxis using extracellular matrix as a migratory cue used only by mesenchymal
cells that have strong substrate adhesion? Do low-​adhesion amoeboid cells still sense
these cues?
Durotaxis
How do cells sense durotactic cues? Are there multiple/cell type-​specific ‘receptors’
for durotaxis, and how do they connect to the cell polarity and translocation machinery?
Topotaxis
How are topotactic cues sensed by cells? Are there different mechanisms that are
specific to particular 2D or 3D geometries? How does topology sensing by membrane
curvature-​sensitive proteins affect leading-​edge protrusions and translate into guided
migration?
Galvanotaxis
Electrophoretic migration of cell surface receptors within the plane of the plasma
membrane and their activation seems to be critical for galvanotaxis, but how are those
receptors activated by electric current-​induced clustering? Is there polarized autocrine
secretion of ligands that accompanies this clustering or is clustering of receptors
sufficient to activate signalling in a ligand-​independent manner?
10 years. New approaches for producing stable and/or
tunable gradients of various factors, controlled substrate
geometries and new methods for imaging cells in 3D,
physiological contexts have expanded our knowledge of
the molecular and cellular processes required to execute
directed migration. The unifying, conceptual framework
presented herein has allowed us to highlight this pro-
gress. Going forwards, we hope that this framework
will prompt the field to fill in gaps in our understanding
(see Box 2).
One of the broader challenges in studying directed
cell migration is understanding it in physiological con-
texts. Studying cell migration on the rigid, 2D surfaces
commonly used for microscopy has been criticized
as not reflecting these physiological contexts222. To be
fair, the tools and imaging approaches needed to study
migration in these more physiological contexts or in
living organisms have only recently become widely
available to researchers, and the field will need to con-
tinue to embrace this transition. However, studying
directed cell migration using simplified 2D systems
and model organisms has revealed underlying princi-
ples and insights that have become the building blocks
to understand migration in more complex systems. By
analogy, studying the biochemical properties of purified
proteins, even though these systems do not replicate the
complexity of the cytoplasm, continues to yield essential
information. Multiple approaches to problems as com-
plex as directed migration are required to achieve the
level of understanding necessary to manipulate this
process or develop therapeutic interventions.
A particularly difficult aspect of understanding
directed migration in vivo is that cells are being exposed
to multiple migration-​inducing cues simultaneously.
This ‘multicue’ problem is both fascinating and experi-
mentally challenging. For example, if a cell is presented
with a chemotactic cue orienting its migration in one
direction but a topological or durotactic cue is orienting
it in the opposite direction, how does the cell prioritize
or integrate these conflicting signals? One compelling
recent example of this problem comes from D. melano­
gaster border cell migration where the relative con-
tributions of chemotactic and topological cues were
dissected by elegant genetic approaches223. However,
knowing the specific perturbations necessary to dissect
multicue migration in vivo requires a specific under-
standing of the underlying processes, and more work in
simplified, single-​cue systems will be needed. In a related
problem, how do cells switch their migration direction?
For example, neutrophils initially home to injured sites
but later reverse their migration and move away from
the inflamed area as part of the resolution process224–226.
Again, a more comprehensive understanding of the
mechanisms of directed migration will be required to
understand complex cases such as these.
A deeper understanding of directed cell migration
will be necessary to effectively treat human diseases.
Interventions that could block tumour cells from leaving
the primary tumour or intravasating into blood or lymph
vessels could be a potent way to block metastasis —
the clinically most deadly aspect of cancer. Similarly,
attenuating the recruitment of immune cells to inflamed
areas in the context of chronic illnesses such as arthritis,
asthma and atherosclerosis could improve disease out-
come. Conversely, encouraging the migration of subsets
of immune cells into tumours could improve antican-
cer immune therapies. The inappropriate migration of
immune cells or fibroblasts can also lead to autoimmune
disorders and fibrosis, respectively. Finally, finding ways
for neuronal cells to ignore ‘stop’ cues such as in nerve
injuries could lead to regenerative treatments for paraly­
sis. Understanding how cells, using different migration
modes (for example, amoeboid versus mesenchymal),
migrate in response to the various chemical, mecha­
nical, geometric and electrical cues that are present
in these disease states has the potential to lead to the
identification of novel therapeutic avenues.
Published online 14 May 2021

1. Borrell, V. Recent advances in understanding
neocortical development. F1000Res https://doi.org/
10.12688/f1000research.20332.1 (2019).
2. Pawar, K. B., Desai, S., Bhonde, R. R., Bhole, R. P.
& Deshmukh, A. A. Wound with diabetes:
present scenario and future. Curr. Diabetes Rev.
https://doi.org/10.2174/157339981666620070318
0137 (2020).
3. Janssen, E. & Geha, R. S. Primary immunodeficiencies
caused by mutations in actin regulatory proteins.
Immunol. Rev. 287, 121–134 (2019).
4. Novikov, N. M., Zolotaryova, S. Y., Gautreau, A. M.
& Denisov, E. V. Mutational drivers of cancer cell
migration and invasion. Br. J. Cancer https://doi.org/
10.1038/s41416-020-01149-0 (2020).
5. García-​Cuesta, E. M. et al. The role of the CXCL12/
CXCR4/ACKR3 axis in autoimmune diseases.
Front. Endocrinol. 10, 585 (2019).
6. Glass, D. S. et al. Idiopathic pulmonary fibrosis:
molecular mechanisms and potential treatment
approaches. Respir Investig. 58, 320–335
(2020).
7. Abercrombie, M. The Croonian Lecture,
1978 - the crawling movement of metazoan cells.
Proc. Royal Soc. B Biol. Sci. 207, 129–147
(1980).
8. Abercrombie, M., Dunn, G. A. & Heath, J. P.
The shape and movement of fibroblasts in culture.
Soc. Gen. Physiol. Ser. 32, 57–70 (1977).
9. Sardelli, L. et al. Engineering biological gradients.
J. Appl. Biomater. Funct. Mater. 17,
2280800019829023 (2019).
10. Buttenschön, A. & Edelstein-​Keshet, L. Bridging from
single to collective cell migration: a review of models
and links to experiments. PLoS Comput. Biol. 16,
e1008411 (2020).
11. Wilkinson, P. C. Random locomotion; chemotaxis
and chemokinesis. a guide to terms defining cell
locomotion. Immunol. Today 6, 273–278 (1985).

NAture RevIews | MolECular CEll BioloGy volume 22 | August 2021 | 543

0123456789();:
Reviews
12. Petrie, R. J., Doyle, A. D. & Yamada, K. M. Random
versus directionally persistent cell migration.
Nat. Rev. Mol. Cell Biol. 10, 538–549 (2009).
13. Dorward, D. A. et al. The role of formylated peptides
and formyl peptide receptor 1 in governing neutrophil
function during acute inflammation. Am. J. Pathol.
185, 1172–1184 (2015).
14. Ehrengruber, M. U., Geiser, T. & Deranleau, D. A.
Activation of human neutrophils by C3a and C5A.
Comparison of the effects on shape changes,
chemotaxis, secretion, and respiratory burst.
FEBS Lett. 346, 181–184 (1994).
15. Sadik, C. D. & Luster, A. D. Lipid-​cytokine-chemokine
cascades orchestrate leukocyte recruitment in
inflammation. J. Leukoc. Biol. 91, 207–215 (2012).
16. McDonald, B. & Kubes, P. Cellular and molecular
choreography of neutrophil recruitment to sites of
sterile inflammation. J. Mol. Med. 89, 1079–1088
(2011).
17. Junger, W. G. Immune cell regulation by autocrine
purinergic signalling. Nat. Rev. Immunol. 11,
201–212 (2011).
18. Yoo, S. K., Starnes, T. W., Deng, Q. & Huttenlocher, A.
Lyn is a redox sensor that mediates leukocyte wound
attraction in vivo. Nature 480, 109–112 (2011).
19. Katikaneni, A. et al. Lipid peroxidation regulates
long-​range wound detection through 5-lipoxygenase
in zebrafish. Nat. Cell Biol. 22, 1049–1055 (2020).
20. Blockus, H. & Chédotal, A. Slit-​Robo signaling.
Development 143, 3037–3044 (2016).
21. Boyer, N. P. & Gupton, S. L. Revisiting netrin-1: one
who guides (axons). Front. Cell Neurosci. 12, 221
(2018).
22. Hu, S. & Zhu, L. Semaphorins and their receptors:
from axonal guidance to atherosclerosis. Front.
Physiol. 9, 1236 (2018).
23. Kania, A. & Klein, R. Mechanisms of ephrin-​Eph
signalling in development, physiology and disease.
Nat. Rev. Mol. Cell Biol. 17, 240–256 (2016).
24. Crick, F. Diffusion in embryogenesis. Nature 225,
420–422 (1970).
25. Majumdar, R., Sixt, M. & Parent, C. A. New paradigms
in the establishment and maintenance of gradients
during directed cell migration. Curr. Opin. Cell Biol.
30, 33–40 (2014).
26. Lau, S. et al. A negative-​feedback loop maintains
optimal chemokine concentrations for directional cell
migration. Nat. Cell Biol. 22, 266–273 (2020).
27. Stapornwongkul, K. S., de Gennes, M., Cocconi, L.,
Salbreux, G. & Vincent, J. P. Patterning and growth
control in vivo by an engineered GFP gradient. Science
370, 321–327 (2020).
Using a synthetic morphogen in D. melanogaster,
the authors show that the expression of non-​
signalling decoy receptors improves the ability of
receptors engineered to respond to the synthetic
morphogen to organize patterning and growth
in vivo.
28. Tweedy, L., Knecht, D. A., Mackay, G. M. & Insall, R. H.
Self-​generated chemoattractant gradients: attractant
depletion extends the range and robustness of
chemotaxis. PLoS Biol. 14, e1002404 (2016).
29. Muinonen-​Martin, A. J. et al. Melanoma cells break
down LPA to establish local gradients that drive
chemotactic dispersal. PLoS Biol. 12, e1001966
(2014).
30. Juin, A. et al. N-​WASP control of LPAR1 trafficking
establishes response to self-​generated LPA gradients
to promote pancreatic cancer cell metastasis. Dev. Cell
https://doi.org/10.1016/j.devcel.2019.09.018 (2019).
31. Tweedy, L. et al. Seeing around corners: cells solve
mazes and respond at a distance using attractant
breakdown. Science https://doi.org/10.1126/science.
aay9792 (2020).
32. Pond, K. W., Doubrovinski, K. & Thorne, C. A.
Wnt/β-catenin signaling in tissue self-​organization.
Genes (Basel) https://doi.org/10.3390/
genes11080939 (2020).
33. Lu, P., Takai, K., Weaver, V. M. & Werb, Z. Extracellular
matrix degradation and remodeling in development
and disease. Cold Spring Harb. Perspect Biol. https://
doi.org/10.1101/cshperspect.a005058 (2011).
34. Afonso, P. V., McCann, C. P., Kapnick, S. M. &
Parent, C. A. Discoidin domain receptor 2 regulates
neutrophil chemotaxis in 3D collagen matrices. Blood
121, 1644–1650 (2013).
35. Shields, J. D. et al. Autologous chemotaxis as a
mechanism of tumor cell homing to lymphatics via
interstitial flow and autocrine CCR7 signaling. Cancer
Cell 11, 526–538 (2007).
36. Polacheck, W. J., Charest, J. L. & Kamm, R. D.
Interstitial flow influences direction of tumor cell
544 | August 2021 | volume 22
migration through competing mechanisms. Proc. Natl
Acad. Sci. USA 108, 11115–11120 (2011).
37. Müller, P., Rogers, K. W., Yu, S. R., Brand, M. &
Schier, A. F. Morphogen transport. Development 140,
1621–1638 (2013).
38. Restrepo, S., Zartman, J. J. & Basler, K. Coordination
of patterning and growth by the morphogen DPP.
Curr. Biol. 24, R245–R255 (2014).
39. Kriebel, P. W., Barr, V. A., Rericha, E. C., Zhang, G.
& Parent, C. A. Collective cell migration requires
vesicular trafficking for chemoattractant delivery at
the trailing edge. J. Cell Biol. 183, 949–961 (2008).
40. Kriebel, P. W. et al. Extracellular vesicles direct
migration by synthesizing and releasing chemotactic
signals. J. Cell Biol. 217, 2891–2910 (2018).
41. Esser, J. et al. Exosomes from human macrophages
and dendritic cells contain enzymes for leukotriene
biosynthesis and promote granulocyte migration.
J. Allergy Clin. Immunol. 126, 1032–1040 (2010).
42. Afonso, P. V. et al. LTB4 is a signal-​relay molecule
during neutrophil chemotaxis. Dev. Cell 22,
1079–1091 (2012).
43. Lämmermann, T. et al. Neutrophil swarms require
LTB4 and integrins at sites of cell death in vivo. Nature
498, 371–375 (2013).
44. Brown, M. et al. Lymphatic exosomes promote
dendritic cell migration along guidance cues.
J. Cell. Biol. 217, 2205–2221 (2018).
45. Chen, T., Guo, J., Yang, M., Zhu, X. & Cao, X.
Chemokine-​containing exosomes are released from
heat-​stressed tumor cells via lipid raft-​dependent
pathway and act as efficient tumor vaccine.
J. Immunol. 186, 2219–2228 (2011).
46. Lim, K. et al. Neutrophil trails guide influenza-​specific
CD8+ T cells in the airways. Science 349, aaa4352
(2015).
47. Sung, B. H., Ketova, T., Hoshino, D., Zijlstra, A. &
Weaver, A. M. Directional cell movement through
tissues is controlled by exosome secretion.
Nat. Commun. 6, 7164 (2015).
48. Sung, B. H. & Weaver, A. M. Exosome secretion
promotes chemotaxis of cancer cells. Cell Adh. Migr.
11, 187–195 (2017).
49. Hynes, R. O. Stretching the boundaries of extracellular
matrix research. Nat. Rev. Mol. Cell Biol. 15,
761–763 (2014).
50. Charras, G. & Sahai, E. Physical influences of the
extracellular environment on cell migration.
Nat. Rev. Mol. Cell Biol. 15, 813–824 (2014).
51. Mouw, J. K., Ou, G. & Weaver, V. M. Extracellular
matrix assembly: a multiscale deconstruction.
Nat. Rev. Mol. Cell Biol. 15, 771–785 (2014).
52. Bonnans, C., Chou, J. & Werb, Z. Remodelling the
extracellular matrix in development and disease.
Nat. Rev. Mol. Cell Biol. 15, 786–801 (2014).
53. Albacete-​Albacete, L. et al. ECM deposition is driven
by caveolin-1-dependent regulation of exosomal
biogenesis and cargo sorting. J. Cell Biol. https://doi.
org/10.1083/jcb.202006178 (2020).
54. Zimmerman, S. P., Asokan, S. B., Kuhlman, B. &
Bear, J. E. Cells lay their own tracks - optogenetic
Cdc42 activation stimulates fibronectin deposition
supporting directed migration. J. Cell Sci. 130,
2971–2983 (2017).
55. Singh, P., Carraher, C. & Schwarzbauer, J. E. Assembly
of fibronectin extracellular matrix. Annu. Rev. Cell Dev.
Biol. 26, 397–419 (2010).
56. Miller, C. G., Budoff, G., Prenner, J. L. &
Schwarzbauer, J. E. Minireview: fibronectin in retinal
disease. Exp. Biol. Med. 242, 1–7 (2017).
57. Schultz, G. S. & Wysocki, A. Interactions between
extracellular matrix and growth factors in wound
healing. Wound Repair Regen. 17, 153–162 (2009).
58. Li, Q., Park, P. W., Wilson, C. L. & Parks, W. C.
Matrilysin shedding of syndecan-1 regulates
chemokine mobilization and transepithelial efflux of
neutrophils in acute lung injury. Cell 111, 635–646
(2002).
59. Schumann, K. et al. Immobilized chemokine fields
and soluble chemokine gradients cooperatively shape
migration patterns of dendritic cells. Immunity 32,
703–713 (2010).
60. Zhu, M. et al. Spatial mapping of tissue properties
in vivo reveals a 3D stiffness gradient in the mouse
limb bud. Proc. Natl Acad. Sci. USA 117, 4781–4791
(2020).
This is a technical tour de force demonstrating
the existence of stiffness gradients in mouse
embryos.
61. Tran, V. D. & Kumar, S. Transduction of cell and matrix
geometric cues by the actin cytoskeleton. Curr. Opin.
Cell Biol. 68, 64–71 (2020).
0123456789();:
62. Cai, L., Xiong, X., Kong, X. & Xie, J. The role of the lysyl
oxidases in tissue repair and remodeling: a concise
review. Tissue Eng. Regen. Med. 14, 15–30 (2017).
63. Handorf, A. M., Zhou, Y., Halanski, M. A. & Li, W. J.
Tissue stiffness dictates development, homeostasis,
and disease progression. Organogenesis 11, 1–15
(2015).
64. Ye, M. et al. Evolving roles of lysyl oxidase family in
tumorigenesis and cancer therapy. Pharmacol. Ther.
215, 107633 (2020).
65. Alcudia, J. F. et al. Lysyl oxidase and endothelial
dysfunction: mechanisms of lysyl oxidase down-​
regulation by pro-​inflammatory cytokines.
Front. Biosci. 13, 2721–2727 (2008).
66. Chen, Y., Peng, W., Raffetto, J. D. & Khalil, R. A.
Matrix metalloproteinases in remodeling of lower
extremity veins and chronic venous disease.
Prog. Mol. Biol. Transl Sci. 147, 267–299 (2017).
67. Quintero-​Fabián, S. et al. Role of matrix
metalloproteinases in angiogenesis and cancer.
Front. Oncol. 9, 1370 (2019).
68. Kaverina, I., Krylyshkina, O. & Small, J. V. Microtubule
targeting of substrate contacts promotes their
relaxation and dissociation. J. Cell. Biol. 146,
1033–1044 (1999).
69. Paul, C. D., Mistriotis, P. & Konstantopoulos, K.
Cancer cell motility: lessons from migration in confined
spaces. Nat. Rev. Cancer 17, 131–140 (2017).
70. Wolf, K., Müller, R., Borgmann, S., Bröcker, E. B.
& Friedl, P. Amoeboid shape change and contact
guidance: T-​lymphocyte crawling through fibrillar
collagen is independent of matrix remodeling by
MMPs and other proteases. Blood 102, 3262–3269
(2003).
71. Renkawitz, J. et al. Nuclear positioning facilitates
amoeboid migration along the path of least resistance.
Nature 568, 546–550 (2019).
This seminal article shows that leukocytes position
the nucleus frontwards to gauge the available
space and choose their path.
72. Castro-​Castro, A. et al. Cellular and molecular
mechanisms of MT1-MMP-​dependent cancer cell
invasion. Annu. Rev. Cell Dev. Biol. 32, 555–576
(2016).
73. Wyckoff, J. B., Pinner, S. E., Gschmeissner, S.,
Condeelis, J. S. & Sahai, E. ROCK- and myosin-​
dependent matrix deformation enables protease-​
independent tumor-​cell invasion in vivo. Curr. Biol. 16,
1515–1523 (2006).
74. Fisher, K. E. et al. MT1-MMP- and Cdc42-dependent
signaling co-​regulate cell invasion and tunnel
formation in 3D collagen matrices. J. Cell Sci. 122,
4558–4569 (2009).
75. Weigelin, B., Bakker, G. J. & Friedl, P. Intravital
third harmonic generation microscopy of collective
melanoma cell invasion: Principles of interface
guidance and microvesicle dynamics. Intravital 1,
32–43 (2012).
76. Du Bois-​Reymond, E. H. Vorläufiger Abriss einer
Untersuchung über den sogenannten Froschstrom und
über die elektromotorischen Fische. Ann. Phys. https://
doi.org/10.1002/andp.18431340102 (1843).
77. Zhao, M. et al. Electrical signals control wound healing
through phosphatidylinositol-3-OH kinase-​gamma and
PTEN. Nature 442, 457–460 (2006).
78. Hotary, K. B. & Robinson, K. R. Endogenous electrical
currents and voltage gradients in Xenopus embryos
and the consequences of their disruption. Dev. Biol.
166, 789–800 (1994).
79. Klein, P. S. et al. A chemoattractant receptor controls
development in Dictyostelium discoideum. Science
241, 1467–1472 (1988).
80. Murphy, P. M. & Tiffany, H. L. Cloning of complementary
DNA encoding a functional human interleukin-8
receptor. Science 253, 1280–1283 (1991).
81. Holmes, W. E., Lee, J., Kuang, W. J., Rice, G. C. &
Wood, W. I. Structure and functional expression
of a human interleukin-8 receptor. Science 253,
1278–1280 (1991).
82. Griffith, J. W., Sokol, C. L. & Luster, A. D. Chemokines
and chemokine receptors: positioning cells for host
defense and immunity. Annu. Rev. Immunol. 32,
659–702 (2014).
83. Hughes, C. E. & Nibbs, R. J. B. A guide to chemokines
and their receptors. FEBS J. 285, 2944–2971 (2018).
84. Zlotnik, A. & Yoshie, O. Chemokines: a new
classification system and their role in immunity.
Immunity 12, 121–127 (2000).
85. Sallusto, F. & Baggiolini, M. Chemokines and
leukocyte traffic. Nat. Immunol. 9, 949–952 (2008).
86. Locati, M. & Murphy, P. M. Chemokines and
chemokine receptors: biology and clinical relevance
www.nature.com/nrm

in inflammation and AIDS. Annu. Rev. Med. 50,
425–440 (1999).
87. Proudfoot, A. E. Chemokine receptors: multifaceted
therapeutic targets. Nat. Rev. Immunol. 2, 106–115
(2002).
88. He, H. Q. & Ye, R. D. The formyl peptide
receptors: diversity of ligands and mechanism for
recognition. Molecules https://doi.org/10.3390/
molecules22030455 (2017).
89. Yokomizo, T. Two distinct leukotriene B4 receptors,
BLT1 and BLT2. J. Biochem. 157, 65–71 (2015).
90. Sadik, C. D., Kim, N. D. & Luster, A. D. Neutrophils
cascading their way to inflammation. Trends Immunol.
32, 452–460 (2011).
91. Anand-​Apte, B. & Zetter, B. Signaling mechanisms in
growth factor-​stimulated cell motility. Stem Cells 15,
259–267 (1997).
92. Vorotnikov, A. V. & Tyurin-​Kuzmin, P. A. Chemotactic
signaling in mesenchymal cells compared to amoeboid
cells. Genes Dis. 1, 162–173 (2014).
93. Bear, J. E. & Haugh, J. M. Directed migration
of mesenchymal cells: where signaling and the
cytoskeleton meet. Curr. Opin. Cell Biol. 30, 74–82
(2014).
94. Gorla, M. & Bashaw, G. J. Molecular mechanisms
regulating axon responsiveness at the midline.
Dev. Biol. 466, 12–21 (2020).
95. Bellon, A. & Mann, F. Keeping up with advances in
axon guidance. Curr. Opin. Neurobiol. 53, 183–191
(2018).
96. Tessier-​Lavigne, M. & Goodman, C. S. The molecular
biology of axon guidance. Science 274, 1123–1133
(1996).
97. Mirakaj, V. & Rosenberger, P. Immunomodulatory
functions of neuronal guidance proteins. Trends
Immunol. 38, 444–456 (2017).
98. Wu, J. Y. et al. The neuronal repellent Slit inhibits
leukocyte chemotaxis induced by chemotactic factors.
Nature 410, 948–952 (2001).
99. Tole, S. et al. The axonal repellent, Slit2, inhibits
directional migration of circulating neutrophils.
J. Leukoc. Biol. 86, 1403–1415 (2009).
100. Pilling, D., Chinea, L. E., Consalvo, K. M. & Gomer, R. H.
Different isoforms of the neuronal guidance molecule
Slit2 directly cause chemoattraction or chemorepulsion
of human neutrophils. J. Immunol. 202, 239–248
(2019).
101. Huttenlocher, A. & Horwitz, A. R. Integrins in cell
migration. Cold Spring Harb. Perspect Biol. 3,
a005074 (2011).
102. Henning Stumpf, B., Ambriovic´-Ristov, A., Radenovic, A.
& Smith, A. S. Recent advances and prospects in the
research of nascent adhesions. Front. Physiol. 11,
574371 (2020).
103. Schwarz, J. et al. Dendritic cells interpret haptotactic
chemokine gradients in a manner governed by signal-​
to-noise ratio and dependent on GRK6. Curr Biol 27,
1314–1325 (2017).
This study shows differential regulation of the
same chemokine receptor in the context of a
chemotactic and haptotactic cue.
104. Shellard, A. & Mayor, R. Durotaxis: the hard path
from in vitro to in vivo. Dev. Cell https://doi.org/
10.1016/j.devcel.2020.11.019 (2020).
105. Sunyer, R. & Trepat, X. Durotaxis. Curr. Biol. 30,
R383–r387 (2020).
106. Janmey, P. A., Fletcher, D. A. & Reinhart-​King, C. A.
Stiffness sensing by cells. Physiol. Rev. 100, 695–724
(2020).
107. Astrof, N. S., Salas, A., Shimaoka, M., Chen, J.
& Springer, T. A. Importance of force linkage in
mechanochemistry of adhesion receptors.
Biochemistry 45, 15020–15028 (2006).
108. Jiang, G., Huang, A. H., Cai, Y., Tanase, M. &
Sheetz, M. P. Rigidity sensing at the leading edge
through alphavbeta3 integrins and RPTPalpha.
Biophys. J. 90, 1804–1809 (2006).
109. Nordenfelt, P. et al. Direction of actin flow dictates
integrin LFA-1 orientation during leukocyte migration.
Nat. Commun. 8, 2047 (2017).
110. Moore, T. I., Aaron, J., Chew, T. L. & Springer, T. A.
Measuring integrin conformational change on the cell
surface with super-​resolution microscopy. Cell Rep.
22, 1903–1912 (2018).
111. Lämmermann, T. et al. Rapid leukocyte migration by
integrin-​independent flowing and squeezing. Nature
453, 51–55 (2008).
112. Hetmanski, J. H. R. et al. Membrane tension
orchestrates rear retraction in matrix-​directed cell
migration. Dev. Cell 51, 460–475.e410 (2019).
This recent study shows the importance of
membrane tension in directed migration.
NAture RevIews | MolECular CEll BioloGy
113. Sitarska, E. & Diz-​Muñoz, A. Pay attention
to membrane tension: mechanobiology of the
cell surface. Curr. Opin. Cell Biol. 66, 11–18
(2020).
114. Baschieri, F. et al. Frustrated endocytosis controls
contractility-​independent mechanotransduction at
clathrin-​coated structures. Nat. Commun. 9, 3825
(2018).
115. Douguet, D. & Honoré, E. Mammalian
mechanoelectrical transduction: structure and
function of force-​gated ion channels. Cell 179,
340–354 (2019).
116. Goult, B. T., Yan, J. & Schwartz, M. A. Talin as a
mechanosensitive signaling hub. J. Cell Biol. 217,
3776–3784 (2018).
117. Grashoff, C. et al. Measuring mechanical tension
across vinculin reveals regulation of focal adhesion
dynamics. Nature 466, 263–266 (2010).
This seminal article establishes a tension-​based
Förster resonance energy transfer biosensor using
the focal adhesion protein vinculin.
118. Sawada, Y. et al. Force sensing by mechanical
extension of the Src family kinase substrate p130Cas.
Cell 127, 1015–1026 (2006).
119. MacKay, J. L. & Hammer, D. A. Stiff substrates
enhance monocytic cell capture through E-​selectin but
not P-​selectin. Integr. Biol. 8, 62–72 (2016).
120. Sun, X. et al. Mechanosensing through direct binding
of tensed F-​Actin by LIM domains. Dev. Cell 55,
468–482.e467 (2020).
121. Winkelman, J. D., Anderson, C. A., Suarez, C.,
Kovar, D. R. & Gardel, M. L. Evolutionarily diverse
LIM domain-​containing proteins bind stressed actin
filaments through a conserved mechanism. Proc. Natl
Acad. Sci. USA 117, 25532–25542 (2020).
This article, along with Sun et al. (2020),
shows that LIM domain-​containing proteins
bind differentially to actin filaments based on
filament tension.
122. Rong, Y. et al. The Golgi microtubules regulate single
cell durotaxis. EMBO Rep. https://doi.org/10.15252/
embr.202051094 (2021).
123. Guilluy, C. et al. Isolated nuclei adapt to force and
reveal a mechanotransduction pathway in the nucleus.
Nat. Cell Biol. 16, 376–381 (2014).
124. Arsenovic, P. T. et al. Nesprin-2G, a component of the
nuclear LINC complex, is subject to myosin-​dependent
tension. Biophys. J. 110, 34–43 (2016).
125. Gomes, E. R., Jani, S. & Gundersen, G. G. Nuclear
movement regulated by Cdc42, MRCK, myosin, and
actin flow establishes MTOC polarization in migrating
cells. Cell 121, 451–463 (2005).
126. Petrie, R. J., Koo, H. & Yamada, K. M. Generation
of compartmentalized pressure by a nuclear piston
governs cell motility in a 3D matrix. Science 345,
1062–1065 (2014).
127. Lou, H. Y., Zhao, W., Zeng, Y. & Cui, B. The role of
membrane curvature in nanoscale topography-​induced
intracellular signaling. Acc. Chem. Res. 51,
1046–1053 (2018).
128. Driscoll, M. K., Sun, X., Guven, C., Fourkas, J. T. &
Losert, W. Cellular contact guidance through dynamic
sensing of nanotopography. ACS Nano 8, 3546–3555
(2014).
129. Chen, S. et al. Actin cytoskeleton and focal adhesions
regulate the biased migration of breast cancer cells
on nanoscale asymmetric sawteeth. ACS Nano 13,
1454–1468 (2019).
This article shows that breast cancer cell lines
exhibit different migration phenotypes on
asymmetric sawtooth nanopatterns through the
distinct regulation of actin polymerization waves
and focal adhesion dynamics.
130. Le Maout, E., Lo Vecchio, S., Kumar Korla, P.,
Jinn-​Chyuan Sheu, J. & Riveline, D. Ratchetaxis
in channels: entry point and local asymmetry set
cell directions in confinement. Biophys. J. 119,
1301–1308 (2020).
This article shows how cell direction through 3D
ratchet channels is dictated by spatial asymmetry
in two-​sided confined channels and the entry point
of the all-​sided confined channels.
131. Ramirez-​San Juan, G. R., Oakes, P. W. & Gardel, M. L.
Contact guidance requires spatial control of leading-​
edge protrusion. Mol. Biol. Cell 28, 1043–1053
(2017).
132. Jang, K. J. et al. Two distinct filopodia populations at
the growth cone allow to sense nanotopographical
extracellular matrix cues to guide neurite outgrowth.
PLoS ONE 5, e15966 (2010).
133. Kubow, K. E., Shuklis, V. D., Sales, D. J. & Horwitz, A. R.
Contact guidance persists under myosin inhibition due
0123456789();:
Reviews
to the local alignment of adhesions and individual
protrusions. Sci. Rep. 7, 14380 (2017).
This is a key article showing that cells can sense the
topology of aligned fibres even in the absence of
myosin activity.
134. Lou, H. Y. et al. Membrane curvature underlies actin
reorganization in response to nanoscale surface
topography. Proc. Natl Acad. Sci. USA 116,
23143–23151 (2019).
This is an important article identifying membrane
curvature-​sensing BAR-​family protein FBP17
as the bridge between surface topology and
topology-​induced reorganization of the actin
network.
135. McGregor, A. L., Hsia, C. R. & Lammerding, J. Squish
and squeeze-​the nucleus as a physical barrier during
migration in confined environments. Curr. Opin. Cell
Biol. 40, 32–40 (2016).
136. Lomakin, A. J. et al. The nucleus acts as a ruler
tailoring cell responses to spatial constraints. Science
https://doi.org/10.1126/science.aba2894 (2020).
137. Venturini, V. et al. The nucleus measures shape
changes for cellular proprioception to control dynamic
cell behavior. Science https://doi.org/10.1126/science.
aba2644 (2020).
This article, along with Lomakin et al. (2020),
shows that cells interpret cell shape and control
movement in confined spaces by gauging nuclear
deformation and initiating signalling events at
the nucleus.
138. Zhao, M. Electrical fields in wound healing — an
overriding signal that directs cell migration.
Semin. Cell Dev. Biol. 20, 674–682 (2009).
139. Allen, G. M., Mogilner, A. & Theriot, J. A.
Electrophoresis of cellular membrane components
creates the directional cue guiding keratocyte
galvanotaxis. Curr. Biol. 23, 560–568 (2013).
This is a key article establishing the mechanism
of galvanotaxis.
140. Sarkar, A., Kobylkevich, B. M., Graham, D. M.
& Messerli, M. A. Electromigration of cell surface
macromolecules in DC electric fields during cell
polarization and galvanotaxis. J. Theor. Biol. 478,
58–73 (2019).
141. Lin, B. J. et al. Lipid rafts sense and direct electric
field-​induced migration. Proc. Natl Acad. Sci. USA
114, 8568–8573 (2017).
142. Li, X., Miao, Y., Pal, D. S. & Devreotes, P. N.
Excitable networks controlling cell migration during
development and disease. Semin. Cell. Dev. Biol. 100,
133–142 (2020).
143. Xiao, Z., Zhang, N., Murphy, D. B. & Devreotes, P. N.
Dynamic distribution of chemoattractant receptors
in living cells during chemotaxis and persistent
stimulation. J. Cell Biol. 139, 365–374 (1997).
144. Servant, G., Weiner, O. D., Neptune, E. R., Sedat, J. W.
& Bourne, H. R. Dynamics of a chemoattractant
receptor in living neutrophils during chemotaxis.
Mol. Biol. Cell 10, 1163–1178 (1999).
145. Nieto, M. et al. Polarization of chemokine receptors
to the leading edge during lymphocyte chemotaxis.
J. Exp. Med. 186, 153–158 (1997).
146. Bailly, M. et al. Epidermal growth factor receptor
distribution during chemotactic responses. Mol. Biol.
Cell 11, 3873–3883 (2000).
147. Bergeron, J. J., Di Guglielmo, G. M., Dahan, S.,
Dominguez, M. & Posner, B. I. Spatial and temporal
regulation of receptor tyrosine kinase activation and
intracellular signal transduction. Annu. Rev. Biochem.
85, 573–597 (2016).
148. Callan-​Jones, A. C. & Voituriez, R. Actin flows in
cell migration: from locomotion and polarity to
trajectories. Curr. Opin. Cell Biol. 38, 12–17 (2016).
149. Parent, C. A., Blacklock, B. J., Froehlich, W. M.,
Murphy, D. B. & Devreotes, P. N. G protein signaling
events are activated at the leading edge of
chemotactic cells. Cell 95, 81–91 (1998).
150. Servant, G. et al. Polarization of chemoattractant
receptor signaling during neutrophil chemotaxis.
Science 287, 1037–1040 (2000).
151. Haugh, J. M., Codazzi, F., Teruel, M. & Meyer, T.
Spatial sensing in fibroblasts mediated by 3’
phosphoinositides. J. Cell Biol. 151, 1269–1280
(2000).
152. Artemenko, Y., Lampert, T. J. & Devreotes, P. N.
Moving towards a paradigm: common mechanisms
of chemotactic signaling in Dictyostelium and
mammalian leukocytes. Cell Mol. Life Sci. 71,
3711–3747 (2014).
153. Kunisaki, Y. et al. DOCK2 is a Rac activator that
regulates motility and polarity during neutrophil
chemotaxis. J. Cell Biol. 174, 647–652 (2006).
volume 22 | August 2021 | 545
Reviews
154. Zhao, T. et al. Signaling requirements for translocation
of P-​Rex1, a key Rac2 exchange factor involved
in chemoattractant-​stimulated human neutrophil
function. J. Leukoc. Biol. 81, 1127–1136 (2007).
155. Yan, J. et al. A Gβγ effector, ElmoE, transduces GPCR
signaling to the actin network during chemotaxis.
Dev. Cell 22, 92–103 (2012).
156. Asokan, S. B. et al. Mesenchymal chemotaxis requires
selective inactivation of myosin II at the leading edge
via a noncanonical PLCgamma/PKCalpha pathway.
Dev. Cell 31, 747–760 (2014).
157. Hoeller, O. & Kay, R. R. Chemotaxis in the absence
of PIP3 gradients. CB 17, 813–817 (2007).
158. Liu, L., Das, S., Losert, W. & Parent, C. A. mTORC2
regulates neutrophil chemotaxis in a cAMP- and
RhoA-​dependent fashion. Dev. Cell 19, 845–857
(2010).
159. Chen, M. Y., Long, Y. & Devreotes, P. N. A novel
cytosolic regulator, Pianissimo, is required for
chemoattractant receptor and G protein-​mediated
activation of the 12 transmembrane domain adenylyl
cyclase in Dictyostelium. Genes Dev. 11, 3218–3231
(1997).
160. Lee, S. et al. TOR complex 2 integrates cell movement
during chemotaxis and signal relay in Dictyostelium.
Mol. Biol. Cell 16, 4572–4583 (2005).
161. He, Y. et al. Mammalian target of rapamycin and
Rictor control neutrophil chemotaxis by regulating
Rac/Cdc42 activity and the actin cytoskeleton.
Mol. Biol. Cell 24, 3369–3380 (2013).
162. Chen, L. et al. PLA2 and PI3K/PTEN pathways act
in parallel to mediate chemotaxis. Dev. Cell 12,
603–614 (2007).
163. van Haastert, P. J., Keizer-​Gunnink, I. & Kortholt, A.
Essential role of PI3-kinase and phospholipase A2
in Dictyostelium discoideum chemotaxis. J. Cell Biol.
177, 809–816 (2007).
164. Heit, B. et al. PTEN functions to ‘prioritize’ chemotactic
cues and prevent ‘distraction’ in migrating neutrophils.
Nat. Immunol. 9, 743–752 (2008).
165. Meliton, A. Y. et al. Cytosolic group IVa phospholipase
A2 mediates IL-8/CXCL8-induced transmigration
of human polymorphonuclear leukocytes in vitro.
J. Inflamm. 7, 14 (2010).
166. Yonker, L. M. et al. Neutrophil-​derived cytosolic
PLA2α contributes to bacterial-​induced neutrophil
transepithelial migration. J. Immunol. 199,
2873–2884 (2017).
167. Ma, H., Gamper, M., Parent, C. & Firtel, R. A. The
Dictyostelium MAP kinase kinase DdMEK1 regulates
chemotaxis and is essential for chemoattractant-​
mediated activation of guanylyl cyclase. EMBO J. 16,
4317–4332 (1997).
168. Mendoza, M. C., Booth, E. O., Shaulsky, G. &
Firtel, R. A. MEK1 and protein phosphatase 4
coordinate Dictyostelium development and
chemotaxis. Mol. Cell Biol. 27, 3817–3827 (2007).
169. Heit, B., Tavener, S., Raharjo, E. & Kubes, P.
An intracellular signaling hierarchy determines
direction of migration in opposing chemotactic
gradients. J. Cell Biol. 159, 91–102 (2002).
170. Liu, X. et al. Bidirectional regulation of neutrophil
migration by mitogen-​activated protein kinases.
Nat. Immunol. 13, 457–464 (2012).
171. Mouneimne, G. et al. Spatial and temporal control of
cofilin activity is required for directional sensing during
chemotaxis. Curr. Biol. 16, 2193–2205 (2006).
172. Miao, Y. et al. Altering the threshold of an excitable
signal transduction network changes cell migratory
modes. Nat. Cell Biol. 19, 329–340 (2017).
173. Zhan, H. et al. An excitable Ras/PI3K/ERK
signaling network controls migration and oncogenic
transformation in epithelial cells. Dev. Cell 54,
608–623.e605 (2020).
174. King, S. J. et al. Lamellipodia are crucial for
haptotactic sensing and response. J. Cell Sci. 129,
2329–2342 (2016).
175. Cox, C. D., Bavi, N. & Martinac, B. Biophysical
principles of ion-​channel-mediated mechanosensory
transduction. Cell Rep. 29, 1–12 (2019).
176. Pal, N., Wu, M. & Lu, H. P. Probing conformational
dynamics of an enzymatic active site by an in situ
single fluorogenic probe under piconewton force
manipulation. Proc. Natl Acad. Sci. USA 113,
15006–15011 (2016).
177. Hu, X., Margadant, F. M., Yao, M. & Sheetz, M. P.
Molecular stretching modulates mechanosensing
pathways. Protein Sci. 26, 1337–1351 (2017).
178. Kumar, A. et al. Talin tension sensor reveals novel
features of focal adhesion force transmission and
mechanosensitivity. J. Cell Biol. 213, 371–383
(2016).
546 | August 2021 | volume 22
179. Rothenberg, K. E., Scott, D. W., Christoforou, N.
& Hoffman, B. D. Vinculin force-​sensitive dynamics
at focal adhesions enable effective directed cell
migration. Biophys. J. 114, 1680–1694 (2018).
This recent work using the vinculin Förster
resonance energy transfer-​based tension sensor
shows precise regulation of tension at different
focal adhesions during directed migration.
180. Ringer, P. et al. Multiplexing molecular tension sensors
reveals piconewton force gradient across talin-1.
Nat Methods 14, 1090–1096 (2017).
181. LaCroix, A. S., Lynch, A. D., Berginski, M. E. &
Hoffman, B. D. Tunable molecular tension sensors
reveal extension-​based control of vinculin loading.
eLife https://doi.org/10.7554/eLife.33927 (2018).
182. Huang, D. L., Bax, N. A., Buckley, C. D., Weis, W. I. &
Dunn, A. R. Vinculin forms a directionally asymmetric
catch bond with F-​actin. Science 357, 703–706
(2017).
183. Owen, L. M., Bax, N. A., Weis, W. I. & Dunn, A. R.
The C-​terminal actin binding domain of talin forms
an asymmetric catch bond with F-​actin. Preprint at
bioRxiv https://doi.org/10.1101/2020.09.01.276568
(2020).
184. Singh, A. V. et al. Astrocytes increase ATP exocytosis
mediated calcium signaling in response to microgroove
structures. Sci. Rep. 5, 7847 (2015).
185. Hung, W. C. et al. Confinement sensing and signal
optimization via Piezo1/PKA and myosin II pathways.
Cell Rep. 15, 1430–1441 (2016).
186. Bavi, N., Richardson, J., Heu, C., Martinac, B. &
Poole, K. PIEZO1-mediated currents are modulated
by substrate mechanics. ACS Nano 13, 13545–13559
(2019).
This article shows that the surface geometry,
including roughness and stiffness, modulates
PIEZO1 channel activation in response to the
external mechanical force applied at cell–substrate
interfaces.
187. Subramanian, B. C. et al. The LTB4-BLT1 axis
regulates actomyosin and β2-integrin dynamics during
neutrophil extravasation. J. Cell Biol. https://doi.org/
10.1083/jcb.201910215 (2020).
This article reports a critical role for neutrophil-​
derived LTB4 in the regulation of non-​muscle
myosin II and β2 integrin that depends on the
release of extracellular vesicles during neutrophil
arrest and extravasation in live mice.
188. Gao, R. et al. A large-​scale screen reveals genes that
mediate electrotaxis in Dictyostelium discoideum.
Sci. Signal 8, ra50 (2015).
189. Sánchez, M. F., Els-​Heindl, S., Beck-​Sickinger, A. G.,
Wieneke, R. & Tampé, R. Photo-​induced receptor
confinement drives ligand-​independent GPCR
signaling. Science https://doi.org/10.1126/science.
abb7657 (2021).
190. Smith, H. E. Nematode sperm motility. WormBook
https://doi.org/10.1895/wormbook.1.68.2 (2014).
191. Pollard, T. D. & Borisy, G. G. Cellular motility driven by
assembly and disassembly of actin filaments. Cell 112,
453–465 (2003).
192. Bodor, D. L., Pönisch, W., Endres, R. G. & Paluch, E. K.
Of cell shapes and motion: the physical basis of animal
cell migration. Dev. Cell 52, 550–562 (2020).
193. Krause, M. & Gautreau, A. Steering cell migration:
lamellipodium dynamics and the regulation of
directional persistence. Nat. Rev. Mol. Cell Biol. 15,
577–590 (2014).
194. McCormick, L. E. & Gupton, S. L. Mechanistic
advances in axon pathfinding. Curr. Opin. Cell Biol.
63, 11–19 (2020).
195. Johnson, H. E. et al. F-​actin bundles direct the
initiation and orientation of lamellipodia through
adhesion-​based signaling. J. Cell Biol. 208, 443–455
(2015).
196. Svitkina, T. M. et al. Mechanism of filopodia initiation
by reorganization of a dendritic network. J. Cell Biol.
160, 409–421 (2003).
197. Wu, C. et al. Arp2/3 is critical for lamellipodia
and response to extracellular matrix cues but is
dispensable for chemotaxis. Cell 148, 973–987
(2012).
This is the first article to establish that the Arp2/3
complex is required for ECM haptotaxis but is
dispensable for RTK-​dependent chemotaxis in
fibroblasts.
198. Schaks, M., Giannone, G. & Rottner, K. Actin dynamics
in cell migration. Essays Biochem. 63, 483–495
(2019).
199. Collins, S. R. et al. Using light to shape chemical
gradients for parallel and automated analysis of
chemotaxis. Mol. Syst. Biol. 11, 804 (2015).
0123456789();:
200. Davidson, A. J., Amato, C., Thomason, P. A. &
Insall, R. H. WASP family proteins and formins
compete in pseudopod- and bleb-​based migration.
J. Cell Biol. 217, 701–714 (2018).
201. Vargas, P. et al. Innate control of actin nucleation
determines two distinct migration behaviours in
dendritic cells. Nat. Cell Biol. 18, 43–53 (2016).
202. Menon, S. et al. The E3 ubiquitin ligase TRIM9 is a
filopodia off switch required for netrin-​dependent axon
guidance. Dev. Cell 35, 698–712 (2015).
203. Oakes, P. W. et al. Lamellipodium is a myosin-​
independent mechanosensor. Proc. Natl Acad.
Sci. USA 115, 2646–2651 (2018).
204. Wong, S., Guo, W. H. & Wang, Y. L. Fibroblasts probe
substrate rigidity with filopodia extensions before
occupying an area. Proc. Natl Acad. Sci. USA 111,
17176–17181 (2014).
205. Andrew, N. & Insall, R. H. Chemotaxis in shallow
gradients is mediated independently of PtdIns
3-kinase by biased choices between random
protrusions. Nat. Cell Biol. 9, 193–200 (2007).
206. Welf, E. S., Ahmed, S., Johnson, H. E., Melvin, A. T.
& Haugh, J. M. Migrating fibroblasts reorient
directionality by a metastable, PI3K-​dependent
mechanism. J. Cell Biol. 197, 105–114 (2012).
207. Sun, X. et al. Asymmetric nanotopography biases
cytoskeletal dynamics and promotes unidirectional
cell guidance. Proc. Natl Acad. Sci. USA 112,
12557–12562 (2015).
208. Vicente-​Manzanares, M., Ma, X., Adelstein, R. S. &
Horwitz, A. R. Non-​muscle myosin II takes centre stage
in cell adhesion and migration. Nat. Rev. Mol. Cell
Biol. 10, 778–790 (2009).
209. Plotnikov, S. V., Pasapera, A. M., Sabass, B. &
Waterman, C. M. Force fluctuations within focal
adhesions mediate ECM-​rigidity sensing to guide
directed cell migration. Cell 151, 1513–1527
(2012).
210. Chan, K. T., Bennin, D. A. & Huttenlocher, A.
Regulation of adhesion dynamics by calpain-​mediated
proteolysis of focal adhesion kinase (FAK). J. Biol.
Chem. 285, 11418–11426 (2010).
211. Mayor, R. & Etienne-​Manneville, S. The front and rear
of collective cell migration. Nat. Rev. Mol. Cell Biol.
17, 97–109 (2016).
212. Ladoux, B. & Mège, R. M. Mechanobiology of
collective cell behaviours. Nat. Rev. Mol. Cell Biol. 18,
743–757 (2017).
213. Shellard, A., Szabó, A., Trepat, X. & Mayor, R.
Supracellular contraction at the rear of neural crest
cell groups drives collective chemotaxis. Science 362,
339–343 (2018).
This article shows a rear wheel drive-​like motility
during collective chemotaxis by actomyosin
contractility in a cluster of rear cells.
214. Yam, P. T. et al. Actin-​myosin network reorganization
breaks symmetry at the cell rear to spontaneously
initiate polarized cell motility. J. Cell Biol. 178,
1207–1221 (2007).
215. Tsai, T. Y. et al. Efficient front-​rear coupling in
neutrophil chemotaxis by dynamic myosin II
localization. Dev. Cell 49, 189–205.e186 (2019).
This article shows that the side-​to-side
redistribution of myosin II at the back of cells in
response to leading-​edge turning is critical for
maintaining persistence and turning ability in
neutrophils undergoing chemotaxis.
216. Allen, G. M. et al. Cell mechanics at the rear act to
steer the direction of cell migration. Cell Syst. 11,
286–299.e284 (2020).
217. Charras, G. & Paluch, E. Blebs lead the way: how to
migrate without lamellipodia. Nat. Rev. Mol. Cell Biol.
9, 730–736 (2008).
218. Zatulovskiy, E., Tyson, R., Bretschneider, T. & Kay, R. R.
Bleb-​driven chemotaxis of Dictyostelium cells. J. Cell
Biol. 204, 1027–1044 (2014).
219. Reversat, A. et al. Cellular locomotion using
environmental topography. Nature 582, 582–585
(2020).
This article identifies friction force generated by
retrograde flow of actin propelling cells forward
through a 3D environment in the absence of
integrin engagement.
220. Aoun, L. et al. Amoeboid swimming is propelled by
molecular paddling in lymphocytes. Biophys. J. 119,
1157–1177 (2020).
This fascinating recent work demonstrates a new
mechanism of cell swimming using ‘molecular
paddling’ of cell surface proteins coupled to flow
of the underlying cell cortex.
221. Purcell, E. M. Life at low Reynolds number. Am. J.
Phys. 45, 3–11 (1977).
www.nature.com/nrm

222. Yamada, K. M. & Sixt, M. Mechanisms of 3D cell
migration. Nat. Rev. Mol. Cell Biol. 20, 738–752
(2019).
223. Dai, W. et al. Tissue topography steers migrating
Drosophila border cells. Science 370, 987–990
(2020).
This article shows that while chemical cues promote
posterior movement, topographical cues provide
orthogonal information and a path of least
resistance during the migration of border cells
in the egg chamber of D. melanogaster embryos.
224. Mathias, J. R. et al. Resolution of inflammation by
retrograde chemotaxis of neutrophils in transgenic
zebrafish. J. Leukoc. Biol. 80, 1281–1288 (2006).
225. Wang, J. et al. Visualizing the function and fate of
neutrophils in sterile injury and repair. Science 358,
111–116 (2017).
226. Hind, L. E. & Huttenlocher, A. Neutrophil reverse
migration and a chemokinetic resolution. Dev. Cell 47,
404–405 (2018).
227. Paluch, E. K., Aspalter, I. M. & Sixt, M. Focal
adhesion-​independent cell migration. Annu. Rev. Cell
Dev. Biol. 32, 469–490 (2016).
228. Shellard, A. & Mayor, R. All roads lead to directional
cell migration. Trends Cell Biol. 30, 852–868 (2020).
NAture RevIews | MolECular CEll BioloGy
229. Haeger, A., Wolf, K., Zegers, M. M. & Friedl, P.
Collective cell migration: guidance principles
and hierarchies. Trends Cell Biol. 25, 556–566
(2015).
230. Lämmermann, T. & Sixt, M. Mechanical modes of
‘amoeboid’ cell migration. Curr. Opin. Cell Biol. 21,
636–644 (2009).
231. Pagès, D.-L. et al. Cell clusters adopt a
collective amoeboid mode of migration in
confined non-​adhesive environments. Preprint at
bioRxiv https://doi.org/10.1101/2020.05.28.106203
(2020).
232. Liu, Y. J. et al. Confinement and low adhesion induce
fast amoeboid migration of slow mesenchymal cells.
Cell 160, 659–672 (2015).
233. Ruprecht, V. et al. Cortical contractility triggers a
stochastic switch to fast amoeboid cell motility.
Cell 160, 673–685 (2015).
234. Graziano, B. R. et al. Cell confinement reveals a
branched-​actin independent circuit for neutrophil
polarity. PLoS Biol. 17, e3000457 (2019).
235. Lehmann, S. et al. Hypoxia induces a HIF-1-dependent
transition from collective-​to-amoeboid dissemination
in epithelial cancer cells. Curr. Biol. 27, 392–400
(2017).
0123456789();:
Reviews
Acknowledgements
The authors apologize to their colleagues whose work on this
vast topic they were unable to cite due to length constraints.
They thank M. Butler for designing some of the artwork in
Fig. 3c, and J. Haugh for thoughtful discussions. J.E.B.
acknowledges support from the NIH (R35GM130312 and
U01EB018816). C.A.P. acknowledges support from the NIH
(R01AI152517).
Author contributions
The authors contributed equally to all aspects of the article.
Competing interests
The authors declare no competing interests.
Peer review information
Nature Reviews Molecular Cell Biology thanks R. Insall, M.
Sixt and the other, anonymous, reviewer(s) for their contribution
to the peer review of this work.
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional
claims in published maps and institutional affiliations.
© Springer Nature Limited 2021
volume 22 | August 2021 | 547